Tertiary amine substituted peptides useful as inhibitors of HCV replication

ABSTRACT

The present invention provides tertiary amine substituted peptides of Formula (I) useful as inhibitors of HCV replication. The variables R and R 1 -R 12  in Formula I are described herein. The invention also includes methods for preparing such compounds. The present invention further includes pharmaceutical compositions containing tertiary amine substituted peptides and methods for using such compounds, including methods for using the compounds to treat hepatitis C infection.

This application is a U.S. National Stage application filed under 35U.S.C. §371 from PCT/US2008/002524 filed 26 Feb. 2008 which is a PCTapplication of U.S. Provisional 60/891,634, filed Feb. 26, 2007, whichis hereby incorporated by reference in its entirety.

FIELD OF INVENTION

The present invention provides tertiary amine substituted peptidesuseful as inhibitors of HCV replication. The invention also includesmethods for preparing such compounds. The present invention furtherincludes pharmaceutical compositions containing tertiary aminesubstituted peptides and methods for using such compounds, includingmethods for using the compounds to treat hepatitis C infection.

BACKGROUND

An estimated 3% of the world's population is infected with the hepatitisC virus. Of those exposed to HCV, 80% become chronically infected, atleast 30% develop cirrhosis of the liver and 1-4% develop hepatocellularcarcinoma. Hepatitis C Virus (HCV) is one of the most prevalent causesof chronic liver disease in the United States, reportedly accounting forabout 15 percent of acute viral hepatitis, 60 to 70 percent of chronichepatitis, and up to 50 percent of cirrhosis, end-stage liver disease,and liver cancer. Chronic HCV infection is the most common cause ofliver transplantation in the U.S., Australia, and most of Europe.Hepatitis C causes an estimated 10,000 to 12,000 deaths annually in theUnited States. While the acute phase of HCV infection is usuallyassociated with mild symptoms, some evidence suggests that only about15% to 20% of infected people will clear HCV.

HCV is an enveloped, single-stranded RNA virus that contains apositive-stranded genome of about 9.6 kb. HCV is classified as a memberof the Hepacivirus genus of the family Flaviviridae. At least 4 strainsof HCV, GT-1-GT-4, have been characterized.

The HCV lifecycle includes entry into host cells; translation of the HCVgenome, polyprotein processing, and replicase complex assembly; RNAreplication, and virion assembly and release. Translation of the HCV RNAgenome yields a more than 3000 amino acid long polyprotein that isprocessed by at least two cellular and two viral proteases. The HCVpolyprotein is:

NH2-C-E1-E2-p7-NS2-NS3-NS4A-NS4B—NS5A-NS5B—COOH.

The cellular signal peptidase and signal peptide peptidase have beenreported to be responsible for cleavage of the N-terminal third of thepolyprotein (C-E1-E2-p7) from the nonstructural proteins(NS2-NS3-NS4A-NS4B—NS5A-NS5B). The NS2-NS3 protease mediates a first ciscleavage at the NS2-NS3 site. The NS3-NS4A protease then mediates asecond cis-cleavage at the NS3-NS4A junction. The NS3-NS4A complex thencleaves at three downstream sites to separate the remainingnonstructural proteins. Accurate processing of the polyprotein isasserted to be essential for forming an active HCV replicase complex.

Once the polyprotein has been cleaved, the replicase complex comprisingat least the NS3-NS5B nonstructural proteins assembles. The replicasecomplex is cytoplasmic and membrane-associated. Major enzymaticactivities in the replicase complex include serine protease activity andNTPase helicase activity in NS3, and RNA-dependent RNA polymeraseactivity of NS5B. In the RNA replication process, a complementarynegative strand copy of the genomic RNA is produced. The negative strandcopy is used as a template to synthesize additional positive strandgenomic RNAs that may participate in translation, replication,packaging, or any combination thereof to produce progeny virus. Assemblyof a functional replicase complex has been described as a component ofthe HCV replication mechanism. Provisional application 60/669,872“Pharmaceutical Compositions and Methods of Inhibiting HCV Replication”filed Apr. 11, 2005, is hereby incorporated by reference in its entiretyfor its disclosure related to assembly of the replicase complex.

Current treatment of hepatitis C infection typically includesadministration of an interferon, such as pegylated interferon (IFN), incombination with ribavirin. The success of current therapies as measuredby sustained virologic response (SVR) depends on the strain of HCV withwhich the patient is infected and the patient's adherence to thetreatment regimen. Only 50% of patients infected with HCV strain GT-1exhibit a sustained virological response. Direct acting antiviral agentssuch as VX-950 and NM 283 (prodrug of NM 107) are in clinicaldevelopment for treatment of chronic HCV. Due to lack of effectivetherapies for treatment for certain HCV strains and the high mutationrate of HCV, new therapies are needed. The present invention fulfillsthis need and provides additional advantages, which are describedherein.

SUMMARY OF INVENTION

Compounds useful for treating and preventing hepatitis C infections,pharmaceutical compositions, and methods for use of such compounds areprovided herein.

In one aspect, the present invention includes compounds of Formula I andpharmaceutically acceptable salts thereof.

Within Formula I the variables R and R₁-R₁₂ carry the definitions setforth below.

R is hydroxyl, C₁-C₄alkoxy,

where R₁₃ is defined herein.

R₁ is C₁-C₆alkyl, C₂-C₆alkenyl, C₃-C₇cycloalkyl, phenyl,heterocycloalkyl, or 5- or 6-membered heteroaryl, each of which isoptionally substituted.

R₂ is C₁-C₆ alkyl or C₂-C₆alkenyl; each of which is optionallysubstituted.

Or, R₁ and R₂ are joined to form a 5- to 7-membered heterocycloalkylring containing 0, 1, or 2 additional heteroatoms independently chosenfrom O, N, or S, which ring is substituted with 0 or 1 or moresubstituents independently chosen from halogen, hydroxyl, C₁-C₄alkyl,C₁-C₄alkoxy, oxo, C₁-C₂haloalkyl, and C₁-C₂haloalkoxy.

R₃ and R₁₁ are independently C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkanoyl, ormono- or di-C₁-C₆alkylamino, (C₃-C₇cycloalkyl)C₀-C₄alkyl,(C₃-C₇cycloalkenyl)C₀-C₄alkyl, (phenyl)C₀-C₂alkyl, or(heterocycloalkyl)C₀-C₄alkyl, each of which is substituted with 0 to 3substituents independently chosen from halogen, hydroxyl, amino, cyano,—CONH₂, —COOH, C₁-C₄alkyl, C₂-C₄alkanoyl, C₁-C₄alkoxy, C₁-C₄alkylthio,mono- and di-C₁-C₄alkylamino, C₁-C₂haloalkyl, and C₁-C₂haloalkoxy.

R₄ and R₁₂ are independently hydrogen, halogen, hydroxyl, amino,C₁-C₆alkyl, C₂-C₆alkenyl, or C₁-C₆alkoxy.

Or, R₃ and R₄ are be joined to form a 3- to 7-membered cycloalkyl ringor 5- to 7-membered heterocycloalkyl ring, each of which is substitutedwith 0 to 2 substituents independently chosen from halogen, hydroxyl,amino, cyano, vinyl, C₁-C₂alkyl, C₁-C₂alkoxy, trifluoromethyl, andtrifluoromethoxy.

Or, R₁₁ and R₁₂ are be joined to form a 3- to 7-membered cycloalkyl ringor 5- to 7-membered heterocycloalkyl ring, each of which is substitutedwith 0 to 2 substituents independently chosen from halogen, hydroxyl,amino, cyano, vinyl, C₁-C₂alkyl, C₁-C₂alkoxy, trifluoromethyl, andtrifluoromethoxy.

Or, R₃ is a C₇-C₁₁ saturated or unsaturated hydrocarbon chain that is(i) covalently bound to R₁₁, where R₁₁ is a methylene group or (ii)covalently bound to a cycloalkyl group formed by R₁₁ and R₁₂ beingjoined to from a 3- to 7-membered cycloalkyl ring.

R₅, R₆, R₇, and R₁₀ are independently hydrogen, halogen, cyano, amino,hydroxyl, C₁-C₂alkyl, C₁-C₂alkoxy, C₁-C₂haloalkyl, or C₁-C₂haloalkoxy.

R₈ is a group of the formula —(CH₂)_(n)Y—Z, where n is 0, 1, or 2, andR₉ is hydrogen, halogen, amino, C₁-C₂alkyl, or C₁-C₂alkoxy; R₈ and R₉are taken together to form an optionally substituted 5- to 7-memberedcycloalkyl ring.

R₁₃ is C₁-C₆alkyl, C₂-C₆alkenyl, each of which is substituted with 0 or1 or more substituents independently chosen from halogen, hydroxyl,amino, cyano, amino, C₁-C₄alkoxy, mono or di-C₁-C₄alkylamino.

Or, R₁₃ is (C₃-C₇cycloalkyl)C₀-C₂alkyl, (aryl)C₀-C₂alkyl, (5- to7-membered heterocycloalkyl)C₀-C₂alkyl, or (heteroaryl)C₀-C₂alkyl, eachof which is substituted with 0 or 1 or more substituents independentlychosen from halogen, hydroxyl, amino, cyano, amino, C₁-C₄alkyl,C₁-C₄alkoxy, mono or di-C₁-C₄alkylamino, C₁-C₂haloalkyl, orC₁-C₂haloalkoxy.

Y is absent, CR₁₄R₁₅, NR₁₆, S, —O—, —O(C═O)(NR₁₆)—, —OC₁₄R₁₅—,NH(C═O)(NR₁₆)—, —NR₁₆(C═O)CR₁₄R₁₅—, NH(S═O)(NR₁₆)—, or —O(C═O)—.

Wherein R₁₄ and R₁₅ are independently hydrogen, hydroxyl, halogen,C₁-C₂alkyl, C₁-C₂alkoxy, C₁-C₂haloalkyl, or C₁-C₂haloalkoxy, and R₁₆ ishydrogen, C₁-C₂alkyl, C₁-C₂haloalkyl, or C₁-C₂haloalkoxy.

Z is C₁-C₆alkyl, C₂-C₆alkenyl, (mono- or bicyclic aryl)C₀-C₂alkyl,(mono- or bicyclic heteroaryl)C₀-C₂alkyl, (C₃-C₇cycloalkyl)C₀-C₂alkyl,(5- or 6-membered heterocycloalkyl)C₀-C₂alkyl, partially unsaturatedbicyclic heterocycle, tricyclic aryl, or tricyclic heteroaryl; each ofwhich Z is substituted with 0 or 1 or more substituents independentlychosen from halogen, hydroxyl, amino, cyano, —CONH₂, —COOH, mono- anddi-C₁-C₄alkylsulfonamide, mono- and di-C₁-C₄alkylcarboxamide,C₁-C₄alkyl, C₂-C₄alkanoyl, C₁-C₄alkoxy, C₁-C₄alkylthio, mono- anddi-C₁-C₄alkylamino, C₁-C₄alkylester, C₁-C₂haloalkyl, andC₁-C₂haloalkoxy, and 0 or 1 (C₃-C₇cycloalkyl)C₀-C₂alkyl,(aryl)C₀-C₂alkyl, (phenyl)C₀-C₂alkoxy, (5- or 6-memberedheteroaryl)C₀-C₂alkyl, (5- or 6-membered heteroaryl)C₀-C₂alkoxy,indanyl, (5- or 6-membered heterocycloalkyl)C₀-C₂alkyl, or 9- or 10membered bicyclic heteroaryl, each of which is substituted with 0, 1, or2 substituents independently chosen from (a) and 0 or 1 substituents(b).

Where (a) is chosen from halogen, hydroxyl, amino, cyano, nitro, —COOH,—CONH₂, NOH, C₁-C₄alkyl, C₁-C₄alkoxy, C₂-C₄alkanoyl, C₁-C₄hydroxyalkyl,mono- and di-C₁-C₄alkylamino, C₁-C₄alkylester, mono- anddi-C₁-C₄alkylsulfonamide, mono- and di-C₁-C₄alkylcarboxamide,C₁-C₂haloalkyl, C₁-C₂haloalkoxy, and (b) is phenyl and 5- or 6-memberedheteroaryl, each of which is substituted with 0 or 1 or moresubstituents independently chosen from halogen, hydroxyl, amino, —CHO,—COOH, —NH(C═O)H, C₁-C₄alkyl, C₁-C₂alkoxy, mono- anddi-(C₁-C₄alkyl)amino, mono-and di-(C_(r) C₄alkyl)carboxamide,C₁-C₄alkylester, (C₁-C₄alkylester)amino, C₁-C₂haloalkyl, andC₁-C₂haloalkoxy.

The invention includes pharmaceutical compositions comprising a compoundof the invention or a salt thereof, containing at least onepharmaceutically acceptable carrier. The invention also includespharmaceutical compositions comprising a compound of the invention andcontaining at least one additional active agent. The invention includespackaged pharmaceutical compositions comprising a compound of theinvention in a container and further comprising instructions for usingthe composition to treat a patient infected with a hepatitis C virus orsusceptible to infection with a hepatitis C virus.

In another aspect the invention provides a method for treating orpreventing hepatitis C infection comprising providing an effectiveamount of a compound or salt of the invention to a patient in need ofsuch treatment or prevention.

A method of inhibiting HCV replication in vivo comprising administeringto a patient infected with HCV a concentration of a compound or salt ofthe invention sufficient to inhibit HCV replicon replication in vitro isalso included in the invention.

These and other aspects of the invention will be more clearly understoodwith reference to the following detailed description, examples andclaims.

DETAILED DESCRIPTION

Terminology

Prior to setting forth the invention in detail, it may be helpful toprovide definitions of certain terms to be used herein. Compounds of thepresent invention are described using standard nomenclature. Unlessdefined otherwise, all technical and scientific terms used herein havethe same meaning as is commonly understood by one of skill in the art towhich this invention belongs.

Formula I includes all subformulae thereof. For example Formula Iincludes compounds of Formulas I-IX and the pharmaceutically acceptablesalts, prodrugs, hydrates, polymorphs, and thereof.

The terms “a” and “an” do not denote a limitation of quantity, butrather denote the presence of at least one of the referenced items. Theterm “or” means “and/or”. The terms “comprising”, “having”, “including”,and “containing” are to be construed as open-ended terms (i.e., meaning“including, but not limited to”). Recitation of ranges of values aremerely intended to serve as a shorthand method of referring individuallyto each separate value falling within the range, unless otherwiseindicated herein, and each separate value is incorporated into thespecification as if it were individually recited herein. The endpointsof all ranges are included within the range and independentlycombinable. All methods described herein can be performed in a suitableorder unless otherwise indicated herein or otherwise clearlycontradicted by context. The use of any and all examples, or exemplarylanguage (e.g., “such as”), is intended merely to better illustrate theinvention and does not pose a limitation on the scope of the inventionunless otherwise claimed. No language in the specification should beconstrued as indicating any non-claimed element as essential to thepractice of the invention as used herein. Unless defined otherwise,technical and scientific terms used herein have the same meaning as iscommonly understood by one of skill in the art to which this inventionbelongs.

An “active agent” means a compound (including a compound of Formula I),element, or mixture that when administered to a patient, alone or incombination with another compound, element, or mixture, confers,directly or indirectly, a physiological effect on the patient. Theindirect physiological effect may occur via a metabolite or otherindirect mechanism. When the active agent is a compound, then salts,solvates (including hydrates) of the free compound, crystalline forms,non-crystalline forms, and any polymorphs of the compound are included.Compounds may contain one or more asymmetric elements such asstereogenic centers, stereogenic axes and the like, e.g., asymmetriccarbon atoms, so that the compounds can exist in differentstereoisomeric forms. These compounds can be, for example, racemates oroptically active forms. For compounds with two or more asymmetricelements, these compounds can additionally be mixtures of diastereomers.For compounds having asymmetric centers, all optical isomers in pureform and mixtures thereof are encompassed. In addition, compounds withcarbon-carbon double bonds may occur in Z- and E-forms, with allisomeric forms of the compounds. In these situations, the singleenantiomers, i.e., optically active forms can be obtained by asymmetricsynthesis, synthesis from optically pure precursors, or by resolution ofthe racemates. Resolution of the racemates can also be accomplished, forexample, by conventional methods such as crystallization in the presenceof a resolving agent, or chromatography, using, for example a chiralHPLC column. All forms are contemplated herein regardless of the methodsused to obtain them.

All forms (for example solvates, optical isomers, enantiomeric forms,polymorphs, free compound and salts) of an active agent may be employedeither alone or in combination.

In certain situations, the compounds of Formula I may contain one ormore asymmetric elements such as stereogenic centers, including chiralcenters, stereogenic axes and the like, e.g. asymmetric carbon atoms, sothat the compounds can exist in different stereoisomeric forms. Thesecompounds can be, for example, racemates or optically active forms. Forcompounds with two or more asymmetric elements, these compounds canadditionally be mixtures of diastereomers. For compounds havingasymmetric centers, it should be understood that all of the opticalisomers and mixtures thereof are encompassed. In addition, compoundswith carbon-carbon double bonds may occur in Z- and E-forms, with allisomeric forms of the compounds being included in the present invention.Formula I includes all chiral forms, stereoisomers, diastereomers, andenantiomers of compounds of Formula I.

The term “chiral” refers to molecules, which have the property ofnon-superimposability of the mirror image partner.

“Stereoisomers” are compounds, which have identical chemicalconstitution, but differ with regard to the arrangement of the atoms orgroups in space.

A “diastereomer” is a stereoisomer with two or more centers of chiralityand whose molecules are not mirror images of one another. Diastereomershave different physical properties, e.g., melting points, boilingpoints, spectral properties, and reactivities. Mixtures of diastereomersmay separate under high resolution analytical procedures such aselectrophoresis, crystallization in the presence of a resolving agent,or chromatography, using, for example a chiral HPLC column.

“Enantiomers” refer to two stereoisomers of a compound, which arenon-superimposable mirror images of one another. A 50:50 mixture ofenantiomers is referred to as a racemic mixture or a racemate, which mayoccur where there has been no stereoselection or stereospecificity in achemical reaction or process.

Stereochemical definitions and conventions used herein generally followS. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984)McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S.,Stereochemistry of Organic Compounds (1994) John Wiley & Sons, Inc., NewYork. Many organic compounds exist in optically active forms, i.e., theyhave the ability to rotate the plane of plane-polarized light. Indescribing an optically active compound, the prefixes D and L or R and Sare used to denote the absolute configuration of the molecule about itschiral center(s). The prefixes d and l or (+) and (−) are employed todesignate the sign of rotation of plane-polarized light by the compound,with (−) or l meaning that the compound is levorotatory. A compoundprefixed with (+) or d is dextrorotatory.

A “racemic mixture” or “racemate” is an equimolar (or 50:50) mixture oftwo enantiomeric species, devoid of optical activity. A racemic mixturemay occur where there has been no stereoselection or stereospecificityin a chemical reaction or process.

Where a compound exists in various tautomeric forms, the invention isnot limited to any one of the specific tautomers, but rather includesall tautomeric forms.

The invention includes compounds of Formula I having all possibleisotopes of atoms occurring in the compounds. Isotopes include thoseatoms having the same atomic number but different mass numbers. By wayof general example, and without limitation, isotopes of hydrogen includetritium and deuterium and isotopes of carbon include ¹¹C, ¹³C, and ¹⁴C.

Certain compounds are described herein using a general formula thatincludes variables, e.g. R, R₁-R₁₂, Y and Z. Unless otherwise specified,each variable within Formula I is defined independently of othervariables. Thus, if a group is said to be substituted, e.g. with 0-2 R*,then said group may be substituted with up to two R* groups and R* ateach occurrence is selected independently from the definition of R*.Also, combinations of substituents and/or variables are permissible onlyif such combinations result in stable compounds.

The term “substituted”, as used herein, means that any one or morehydrogens on the designated atom or group is replaced with a selectionfrom the indicated group, provided that the designated atom's normalvalence is not exceeded. When the substituent is oxo (i.e., ═O), then 2hydrogens on the atom are replaced. When aromatic moieties aresubstituted by an oxo group, the aromatic ring is replaced by thecorresponding partially unsaturated ring. For example a pyridyl groupsubstituted by oxo is a pyridone. Combinations of substituents and/orvariables are permissible only if such combinations result in stablecompounds or useful synthetic intermediates. A stable compound or stablestructure is meant to imply a compound that is sufficiently robust tosurvive isolation from a reaction mixture, and subsequent formulationinto an effective therapeutic agent.

A dash (“-”) that is not between two letters or symbols is used toindicate a point of attachment for a substituent. For example,—(CH₂)C₃-C₇cycloalkyl is attached through carbon of the methylene (CH₂)group.

“Alkyl” includes both branched and straight chain saturated aliphatichydrocarbon groups, having the specified number of carbon atoms,generally from 1 to about 12 carbon atoms. The term C₁-C₆alkyl as usedherein indicates an alkyl group having from 1 to about 6 carbon atoms.When C₀-C_(n) alkyl is used herein in conjunction with another group,for example, (phenyl)C₀-C₄ alkyl, the indicated group, in this casephenyl, is either directly bound by a single covalent bond (C₀), orattached by an alkyl chain having the specified number of carbon atoms,in this case from 1 to about 4 carbon atoms. Examples of alkyl include,but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl,3-methylbutyl, t-butyl, n-pentyl, and sec-pentyl.

“Alkanoyl” is an alkyl group as defined above, attached through a keto(—(C═O)—) bridge. Alkanoyl groups have the indicated number of carbonatoms, with the carbon of the keto group being included in the numberedcarbon atoms. For example a C₂alkanoyl group is an acetyl group havingthe formula CH₃(C═O)—.

“Alkenyl” means straight and branched hydrocarbon chains comprising oneor more unsaturated carbon-carbon bonds, which may occur in any stablepoint along the chain. Alkenyl groups described herein typically havefrom 2 to about 12 carbons atoms. Preferred alkenyl groups are loweralkenyl groups, those alkenyl groups having from 2 to about 8 carbonatoms, e.g. C₂-C₈, C₂-C₆, and C₂-C₄ alkenyl groups. Examples of alkenylgroups include ethenyl, propenyl, and butenyl groups.

“Alkoxy” means an alkyl group, as defined above, with the indicatednumber of carbon atoms attached via an oxygen bridge. Examples of alkoxyinclude, but are not limited to, methoxy, ethoxy, 3-hexoxy, and3-methylpentoxy.

“Mono- and/or di-alkylamino” indicates secondary or tertiary alkyl aminogroups, wherein the alkyl groups are as defined above and have theindicated number of carbon atoms. The point of attachment of thealkylamino group is on the nitrogen. The alkyl groups are independentlychosen. Examples of mono- and di-alkylamino groups include ethylamino,dimethylamino, and methyl-propyl-amino. “Mono- and/or dialkylaminoalkyl”groups are mono- and/or di-alkylamino groups attached through an alkyllinker having the specified number of carbon atoms, for example adi-methylaminoethyl group. Tertiary amino substituents may by designatedby nomenclature of the form N—R—N—R′, indicating that the groups R andR′ are both attached to a single nitrogen atom.

“Alkylester” indicates an alkyl group as defined above attached throughan ester linkage. The ester linkage may be in either orientation, e.g. agroup of the formula —O(C═O)alkyl or a group of the formula—(C═O)Oalkyl.

“Alkylthio” indicates an alkyl group as defined above attached through asulfur linkage, i.e. a group of the formula alkyl-S—. Examples includeethylthio and pentylthio.

“Aryl” means aromatic groups containing only carbon in the aromatic ringor rings. Typical aryl groups contain 1 to 3 separate, fused, or pendantrings and from 6 to about 18 ring atoms, without heteroatoms as ringmembers. When indicated, such aryl groups may be further substitutedwith carbon or non-carbon atoms or groups. Bicyclic aryl groups maycontain two fused aromatic rings (naphthyl) or an aromatic ring fused toa 5- to 7-membered non-aromatic cyclic group that optionally contains 1or 2 heteroatoms independently chosen from N, O, and S, for example, a3,4-methylenedioxy-phenyl group. Aryl groups include, for example,phenyl, naphthyl, including 1-naphthyl and 2-naphthyl, and bi-phenyl.

“Mono- and/or di-alkylcarboxamide” indicates groups of formula(alkyl₁)—NH—(C═O)— and (alkyl₁)(alkyl₂)—N—(C═O)— in which the alkyl_(s)and alkyl₂ groups are independently chosen alkyl groups as defined abovehaving the indicated number of carbon atoms. Mono and/ordi-alkylcarboxamide also refers to groups of the formula—NH(C═O)(alkyl₁) and

—N(alkyl₂)(C═O)(alkyl₁), carboxamide groups in which the point ofattachment is the nitrogen atom, in which the alkyl₁ and alkyl₂ groupsare independently chosen alkyl groups as defined above having theindicated number of carbon atoms.

“Mono- and di-alkylsulfonamide” means groups of the formula(alkyl₁)—NH—(SO₂)— and (alkyl₁)(alkyl₂)—N—(SO₂)— in which the alkyl₁ andalkyl₂ groups are independently chosen alkyl groups as defined abovehaving the indicated number of carbon atoms. Mono and/ordi-alkylcarboxamide also refers to groups of the formula—NH(SO₂)(alkyl₁) and —N(alkyl₂)(SO₂)(alkyl₁), sulfonamide groups inwhich the point of attachment is the nitrogen atom, in which the alkyl₁and alkyl₂ groups are independently chosen alkyl groups as defined abovehaving the indicated number of carbon atoms.

“Cycloalkyl” indicates a saturated hydrocarbon ring group, having thespecified number of carbon atoms, usually from 3 to about 8 ring carbonatoms, or from 3 to about 7 carbon atoms. Examples of cycloalkyl groupsinclude cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl as well asbridged or caged saturated ring groups such as norborane or adamantane.(Cycloalkyl)C₀-C₂alkyl means a cycloalkyl group as defined aboveattached to the core group it substitutes via a single covalent bond(C₀alkyl) or via a methylene or ethylene linker.

“Cycloalkenyl” as used herein, means an unsaturated, but not aromatic,hydrocarbon ring having at least one carbon-carbon double bond.Cycloalkenyl groups contain from 4 to about 8 carbon atoms, usually from4 to about 7 carbon atoms. Examples include cyclohexenyl andcyclobutenyl. In the term “(cycloalkenyl)alkyl” the terms “cycloalkenyl”and “alkyl” are as defined above, and the point of attachment is on thealkyl group.

“Mono- or bicyclic heteroaryl” indicates a stable 5- to 7-memberedmonocyclic or 7- to 10-membered bicyclic heterocyclic ring whichcontains at least 1 aromatic ring that contains from 1 to 4, orpreferably from 1 to 3, heteroatoms chosen from N, O, and S, withremaining ring atoms being carbon. When the total number of S and Oatoms in the heteroaryl group exceeds 1, these heteroatoms are notadjacent to one another. It is preferred that the total number of S andO atoms in the heteroaryl group is not more than 2. It is particularlypreferred that the total number of S and O atoms in the heteroaryl groupis not more than 1. A nitrogen atom in a heteroaryl group may optionallybe quaternized. When indicated, such heteroaryl groups may be furthersubstituted with carbon or non-carbon atoms or groups. Such substitutionmay include fusion to a 5 to 7-membered saturated cyclic group thatoptionally contains 1 or 2 heteroatoms independently chosen from N, O,and S, to form, for example, a [1,3]dioxolo[4,5-c]pyridyl group. Incertain embodiments 5- to 6-membered heteroaryl groups are preferred.Examples of heteroaryl groups include, but are not limited to, pyridyl,indolyl, pyrimidinyl, pyridizinyl, pyrazinyl, imidazolyl, oxazolyl,furanyl, thiophenyl, thiazolyl, triazolyl, tetrazolyl, isoxazolyl,quinolinyl, pyrrolyl, pyrazolyl, benz[b]thiophenyl, isoquinolinyl,quinazolinyl, quinoxalinyl, thienyl, isoindolyl, and5,6,7,8-tetrahydroisoquinoline.

“Heterocycloalkyl” means a saturated cyclic group containing from 1 toabout 3 heteroatoms chosen from N, O, and S, with remaining ring atomsbeing carbon. Heterocycloalkyl groups have from 3 to about 8 ring atoms,and more typically have from 5 to 7 ring atoms. Examples ofheterocycloalkyl groups include morpholinyl, piperazinyl, piperidinyl,and pyrrolidinyl groups. A nitrogen in a heterocycloalkyl group mayoptionally be quaternized. A (heterocycloalkyl)C₀-C₂alkyl means aheterocycloalkyl group as defined above attached to the core group itsubstitutes via a single covalent bond (C₀alkyl) or via a methylene orethylene linker.

“Haloalkyl” indicates both branched and straight-chain alkyl groupshaving the specified number of carbon atoms, substituted with 1 or morehalogen atoms, generally up to the maximum allowable number of halogenatoms. Examples of haloalkyl include, but are not limited to,trifluoromethyl, difluoromethyl, 2-fluoroethyl, and penta-fluoroethyl.

“Haloalkoxy” indicates a haloalkyl group as defined above attachedthrough an oxygen bridge (oxygen of an alcohol radical).

“Halo” or “halogen” as used herein refers to fluoro, chloro, bromo, oriodo.

“Hydroxyalkyl” is an alkyl group as defined herein substituted with atleast on hydroxyl substituent.

The phrase “optionally substituted” indicates that such groups mayeither be unsubstituted or substituted at one or more of any of theavailable positions, typically 1, 2, 3, or 4 positions, by one or moresuitable groups such as those disclosed herein.

The phrase Suitable groups that may be present on an “optionallysubstituted” position include, but are not limited to, e.g., halogen,cyano, hydroxyl, amino, nitro, oxo, azido, alkanoyl (such as a C₂-C₆alkanoyl group such as acyl or the like); carboxamido; alkylcarboxamide;alkylester groups; alkyl groups, alkoxy groups, alkylthio groupsincluding those having one or more thioether linkages, alkylsulfinylgroups including those having one or more sulfinyl linkages,alkylsulfonyl groups including those having one or more sulfonyllinkages, mono- and di-aminoalkyl groups including groups having one ormore N atoms, all of the foregoing optional alkyl substituents may haveone or more methylene group replaced by an oxygen or —NH—, and have fromabout 1 to about 8, from about 1 to about 6, or from 1 to about 4 carbonatoms, cycloalkyl; phenyl; phenylalkyl with benzyl being an exemplaryphenylalkyl group, phenylalkoxy with benzyloxy being an exemplaryphenylalkoxy group; a saturated, unsaturated, or aromatic heterocyclicgroups having 1 ring and one or more N, O or S atoms, e.g. pyridyl,pyrazinyl, pyrimidinyl, furanyl, pyrrolyl, thienyl, thiazolyl,triazinyl, oxazolyl, isoxazolyl, imidazolyl, tetrahydrofuranyl,tetrahydropyranyl, piperidinyl, morpholinyl, piperazinyl, andpyrrolidinyl. Any such groups having additional positions available forsubstitution may be further substituted, e.g with substituentsindependently chosen from, e.g., amino, hydroxyl, alkyl, alkoxy,halogen, haloalkyl, haloalkoxy, and mono- and di-alkylamino.

“Pharmaceutical compositions” are compositions comprising at least oneactive agent, such as a compound or salt of Formula I, and at least oneother substance, such as a carrier, excipient, or diluent.Pharmaceutical compositions meet the U.S. FDA's GMP (good manufacturingpractice) standards for human or non-human drugs.

“Pharmaceutically acceptable salts” includes derivatives of thedisclosed compounds in which the parent compound is modified by makinginorganic and organic, non-toxic, acid or base addition salts thereof.The salts of the present compounds can be synthesized from a parentcompound that contains a basic or acidic moiety by conventional chemicalmethods. Generally, such salts can be prepared by reacting free acidforms of these compounds with a stoichiometric amount of the appropriatebase (such as Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate, or thelike), or by reacting free base forms of these compounds with astoichiometric amount of the appropriate acid. Such reactions aretypically carried out in water or in an organic solvent, or in a mixtureof the two. Generally, non-aqueous media like ether, ethyl acetate,ethanol, isopropanol, or acetonitrile are preferred, where practicable.Salts of the present compounds further include solvates of the compoundsand of the compound salts.

Examples of pharmaceutically acceptable salts include, but are notlimited to, mineral or organic acid salts of basic residues such asamines; alkali or organic salts of acidic residues such as carboxylicacids; and the like. The pharmaceutically acceptable salts include theconventional non-toxic salts and the quaternary ammonium salts of theparent compound formed, for example, from non-toxic inorganic or organicacids. For example, conventional non-toxic acid salts include thosederived from inorganic acids such as hydrochloric, hydrobromic,sulfuric, sulfamic, phosphoric, nitric and the like; and the saltsprepared from organic acids such as acetic, propionic, succinic,glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic,maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic,mesylic, esylic, besylic, sulfanilic, 2-acetoxybenzoic, fumaric,toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic,HOOC—(CH₂)_(n)—COON where n is 0-4, and the like. Lists of additionalsuitable salts may be found, e.g., in Remington's PharmaceuticalSciences, 17th ed., Mack Publishing Company, Easton, Pa., p. 1418(1985).

The term “carrier” applied to pharmaceutical compositions of theinvention refers to a diluent, excipient, or vehicle with which anactive compound is administered.

A “pharmaceutically acceptable excipient” means an excipient that isuseful in preparing a pharmaceutical composition that is generally safe,non-toxic and neither biologically nor otherwise undesirable, andincludes an excipient that is acceptable for veterinary use as well ashuman pharmaceutical use. A “pharmaceutically acceptable excipient” asused in the present application includes both one and more than one suchexcipient.

A “patient” is a human or non-human animal in need of medical treatment.Medical treatment can include treatment of an existing condition, suchas a disease or disorder, prophylactic or preventative treatment, ordiagnostic treatment. In some embodiments the patient is a humanpatient.

“Prodrug” means any compound that becomes compound of the invention whenadministered to a mammalian subject, e.g., upon metabolic processing ofthe prodrug. Examples of prodrugs include, but are not limited to,acetate, formate and benzoate and like derivatives of functional groups(such as alcohol or amine groups) in the compounds of the invention.

“Providing” means giving, administering, selling, distributing,transferring (for profit or not), manufacturing, compounding, ordispensing.

“Providing a compound of Formula I with at least one additional activeagent” means the compound of Formula I and the additional activeagent(s) are provided simultaneously in a single dosage form, providedconcomitantly in separate dosage forms, or provided in separate dosageforms for administration separated by some amount of time that is withinthe time in which both the compound of Formula I and the at least oneadditional active agent are within the blood stream of a patient. Thecompound of Formula I and the additional active agent need not beprescribed for a patient by the same medical care worker. The additionalactive agent or agents need not require a prescription. Administrationof the compound of Formula I or the at least one additional active agentcan occur via any appropriate route, for example, oral tablets, oralcapsules, oral liquids, inhalation, injection, suppositories or topicalcontact.

“Treatment,” as used herein includes providing a compound of Formula Iand at least one additional active agent sufficient to: (a) prevent adisease or a symptom of a disease from occurring in a patient who may bepredisposed to the disease but has not yet been diagnosed as having it(e.g. including diseases that may be associated with or caused by aprimary disease (as in liver fibrosis that can result in the context ofchronic HCV infection); (b) inhibiting the disease, i.e. arresting itsdevelopment; and (c) relieving the disease, i.e., causing regression ofthe disease. “Treating” and “treatment” also means providing atherapeutically effective amount of a compound of Formula I and at leastone additional active agent to a patient having or susceptible to ahepatitis C infection.

A “therapeutically effective amount” of a pharmaceutical combination ofthis invention means an amount effective, when administered to apatient, to provide a therapeutic benefit such as an amelioration ofsymptoms, e.g., an amount effective to decrease the symptoms of ahepatitis C infection. For example a patient infected with a hepatitis Cvirus may present elevated levels of certain liver enzymes, includingAST and ALT. Normal levels of AST are from 5 to 40 units per liter ofserum (the liquid part of the blood) and normal levels of ALT are from 7to 56 units per liter of serum. A therapeutically effect amount is thusan amount sufficient to provide a significant reduction in elevated ASTand ALT levels or an amount sufficient to provide a return of AST andALT levels to the normal range. A therapeutically effective amount isalso an amount sufficient to prevent a significant increase orsignificantly reduce the detectable level of virus or viral antibodiesin the patient's blood, serum, or tissues. One method of determiningtreatment efficacy includes measuring HCV RNA levels by a conventionmethod for determining viral RNA levels such as the Roch TaqMan assay.In certain preferred embodiments treatment reduces HCV RNA levels belowthe limit of quantitation (30 IU/mL, as measured by the Roche TaqMan®assay) or more preferably below the limit of detection (10 IU/mL, RocheTaqMan).

A significant increase or reduction in the detectable level of virus orviral antibodies is any detectable change that is statisticallysignificant in a standard parametric test of statistical significancesuch as Student's T-test, where p<0.05.

Chemical Description

The invention includes compounds of Formula I, as discussed above.Additionally the invention includes, as alternate embodiments, compoundand salts of Formula I in which the variables A and R₃-R₇ carry thevalues set forth below.

The R₁ and R₂ Variables

(a) R₁ is C₁-C₆alkyl, C₂-C₆alkenyl, C₃-C₇cycloalkyl, phenyl,heterocycloalkyl, or 5- or 6-membered heteroaryl, each of which isoptionally substituted; and R₂ is C₁-C₆ alkyl or C₂-C₆alkenyl; each ofwhich is optionally substituted.

(b) R₁ and R₂ are joined to form a 5- to 7-membered heterocycloalkylring, which ring is substituted with 0 or 1 or more substituentsindependently chosen from halogen, hydroxyl, C₁-C₄alkyl, C₁-C₄alkoxy,C₁-C₂haloalkyl, and C₁-C₂haloalkoxy.

(c) R₁ and R₂ are joined to form a pyrrolidinyl, piperidinyl, orpiperazinyl ring, each of which is substituted with 0, 1, or 2substituents independently chosen from halogen, hydroxyl, C₁-C₂alkyl,C₁-C₂alkoxy, trifluoromethyl, and trifluoromethoxy.

The R₃, R₄, R₁₁, and R₁₂ Variables

(d) R₃ is C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkanoyl, or mono- ordi-C₁-C₆alkylamino, (C₃-C₇cycloalkyl)C₀-C₄alkyl,(C₃-C₇cycloalkenyl)C₀-C₄alkyl, (phenyl)C₀-C₂alkyl, or(heterocycloalkyl)C₀-C₄alkyl, each of which is substituted with 0 to 3substituents independently chosen from halogen, hydroxyl, amino, cyano,—CONH₂, —COOH, C₁-C₄alkyl, C₂-C₄alkanoyl, C₁-C₄alkoxy, C₁-C₄alkylthio,mono- and di-C₁-C₄alkylamino, C₁-C₂haloalkyl, and C₁-C₂haloalkoxy, andR₄ is hydrogen, C₁-C₆alkyl, C₂-C₆alkenyl, or C₁-C₆alkoxy.

(e) R₃ is C₁-C₆alkyl, (C₃-C₇cycloalkyl)C₀-C₄alkyl, or(phenyl)C₀-C₂alkyl, each of which is substituted with 0 to 3substituents independently chosen from halogen, hydroxyl, C₁-C₂alkyl,C₁-C₂alkoxy, mono- and di-C₁-C₂alkylamino, C₁-C₂haloalkyl, andC₁-C₂haloalkoxy; and R₄ is hydrogen, C₁-C₂alkyl, or C₁-C₂alkoxy.

(f) R₃ is C₁-C₆alkyl, (C₃-C₇cycloalkyl)C₀-C₄alkyl, or(phenyl)C₀-C₂alkyl.

(g) R₃ is C₁-C₆alkyl.

(h) R₄ is hydrogen.

(i) R₃ and R₄ are be joined to form a cyclopropyl ring, which issubstituted with 0 to 2 substituents independently chosen from vinyl,C₁-C₂alkyl, and C₁-C₂alkoxy.

(j) R₁₁ is C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkanoyl, or mono- ordi-C₁-C₆alkylamino, (C₃-C₇cycloalkyl)C₀-C₄alkyl,(C₃-C₇cycloalkenyl)C₀-C₄alkyl, (phenyl)C₀-C₂alkyl, or(heterocycloalkyl)C₀-C₄alkyl, each of which is substituted with 0 to 3substituents independently chosen from halogen, hydroxyl, amino, cyano,—CONH₂, —COOH, C₁-C₄alkyl, C₂-C₄alkanoyl, C₁-C₄alkoxy, C₁-C₄alkylthio,mono- and di-C₁-C₄alkylamino, C₁-C₂haloalkyl, and C₁-C₂haloalkoxy, andR₁₂ is hydrogen, C₁-C₆alkyl, C₂-C₆alkenyl, or C₁-C₆alkoxy.

(k) R₁₁ is C₁-C₆alkyl, (C₃-C₇cycloalkyl)C₀-C₄alkyl, or(phenyl)C₀-C₂alkyl, each of which is substituted with 0 to 3substituents independently chosen from halogen, hydroxyl, C₁-C₂alkyl,C₁-C₂alkoxy, mono- and di-C₁-C₂alkylamino, C₁-C₂haloalkyl, andC₁-C₂haloalkoxy; and R₁₂ is hydrogen, C₁-C₂alkyl, or C₁-C₂alkoxy.

(l) R₁₁ is C₁-C₆alkyl, (C₃-C₇cycloalkyl)C₀-C₄alkyl, or(phenyl)C₀-C₂alkyl.

(m)_(R) ¹¹ is C₁-C₆alkyl.

(n) R₃ and R₄ are be joined to form a cyclopropyl ring, which issubstituted with 0 to 2 substituents independently chosen from vinyl,C₁-C₂alkyl, and C₁-C₂alkoxy.

(o) The invention includes embodiments in which R₃ and R₁₁ are joined toform a macrocyclic ring. E.g. the invention includes compounds and saltsof Formula II

Within Formula II D is a alkyl or alkenyl group having 6 to 10 carbonatoms.

(p) For example the invention include compounds and salts of Formula M.

(q) The invention includes embodiments in which R₃ is covalently boundto a cycloalkyl group formed by R₁₁ and R₁₂ being joined to from a 3- to7-membered cycloalkyl ring. E.g. the invention includes compounds andsalts of Formula IV.

Within Formula IV D is a alkyl or alkenyl group having 6 to 10 carbonatoms.

(r) For example the invention includes compounds and salt of Formula V.

The R₅, R₆, R₇, and R₁₀ Variables

(s) The invention includes compounds and salts of any of the abovechemical formulae in which R₅, R₆, R₇, and R₁₀ are all hydrogen.

The R₈ and R₉ Variables

(t) R₈ is a group of the formula —(CH₂)_(n)Y—Z, where n is 0, 1, or 2,and R₉ is hydrogen, halogen, amino, C₁-C₂alkyl, or C₁-C₂alkoxy.

(u) n is 0 and Y is —O—, —O(C═O)(NR₁₆)—, —NR₁₆(C═O)—, or —O(C═O)—.

(v) n is 0 and —O— or —O(C═O)—.

(w) Z is a group of the formula

wherein X₁, X₂, X₃, and X₄, are independently N or CH and no more thantwo of X₁-X₄ are N;

R₂₁ represents from 0 to 3 groups independently chosen from halogen,hydroxyl, amino, cyano, —CONH₂, —COOH, C₁-C₄alkyl, C₂-C₄alkanoyl,C₁-C₄alkoxy, C₁-C₄alkylthio, mono- and di-C₁-C₄alkylamino,C₁-C₂haloalkyl, and C₁-C₂haloalkoxy.

R₂₂ is hydrogen, halogen, hydroxyl, amino, cyano, —CONH₂, —COOH,C₂-C₄alkanoyl, C₁-C₄alkoxy, C₁-C₄alkylthio, mono- anddi-C₁-C₄alkylamino, C₁-C₄alkylester, C₁-C₂haloalkyl, andC₁-C₂haloalkoxy, or

R₂₂ is (C₃-C₇cycloalkyl)C₀C₂alkyl, (phenyl)C₀C₂alkyl,(phenyl)C₀C₂alkoxy, (5- or 6-membered heteroaryl)C₀-C₂alkyl, (5- or6-membered heteroaryl)C₀-C₂alkoxy, indanyl, (5- or 6-memberedheterocycloalkyl)C₀C₂alkyl, or 9- or 10 membered bicyclic heteroaryl,each of which is substituted with 0, 1, or 2 substituents independentlychosen from (a) and 0 or 1 substituents (b) where (a) is chosen fromhalogen, hydroxyl, amino, cyano, nitro, —COOH, —CONH₂, CH₃(C═O)NH—,C₁-C₄alkyl, C₁-C₄alkoxy, C₁-C₄hydroxyalkyl, mono- anddi-C₁-C₄alkylamino, C₁-C₄alkylester, mono- and di-C₁-C₄alkylsulfonamide,mono- and di-C₁-C₄alkylcarboxamide, C₁-C₂haloalkyl, and C₁-C₂haloalkoxy,and (b) is phenyl or 5- or 6-membered heteroaryl, each of which issubstituted with 0 or 1 or more of halogen, hydroxyl, C₁-C₄alkyl, andC₁-C₂alkoxy.

(x) Z is a group as defined above in (w) and R₂₂ is (phenyl)C₀C₂alkyl,(phenyl)C₀-C₂alkoxy, or (5- or 6-membered heteroaryl)C₀-C₂alkyl, each ofwhich is substituted with 0, 1, or 2 substituents independently chosenfrom (c) halogen, hydroxyl, amino, cyano, —COOH, —CONH₂, C₁-C₂alkyl,C₁-C₂alkoxy, mono- and di-C₁-C₂alkylamino, trifluoromethyl, andtrifluoromethoxy.

(y) Z is a group as defined above in (w) and R₂₂ is

(z) Z is C₁-C₆alkyl or C₂-C₆alkenyl, each of which Z is substituted with0 or 1 or more substituents independently chosen from halogen, hydroxyl,amino, cyano, —CONH₂, —COOH, mono- and di-C₁-C₄alkylsulfonamide, mono-and di-C₁-C₄alkylcarboxamide, C₁-C₄alkyl, C₂-C₄alkanoyl, C₁-C₄alkoxy,C₁-C₄alkylthio, mono- and di-C₁-C₄alkylamino, C₁-C₄alkylester,C₁-C₂haloalkyl, and C₁-C₂haloalkoxy.

(aa) Z is (phenyl)C₀-C₂alkyl, (naphthyl)C₀-C₂alkyl, (mono- or bicyclicheteroaryl)C₀-C₂alkyl, or tetrahydroquinolinyl, each of which Z issubstituted with 0 or 1 or more substituents independently chosen fromhalogen, hydroxyl, amino, cyano, —CONH₂, —COOH, C₁-C₄alkyl,C₂-C₄alkanoyl, C₁-C₄alkoxy, C₁-C₄alkylthio, mono- anddi-C₁-C₄alkylamino, C₁-C₂haloalkyl, and C₁-C₂haloalkoxy, and 0 or 1C₃-C₇cycloalkyl, aryl, 5- or 6-membered heteroaryl, indanyl, or 5- or6-membered heterocycloalkyl, or 9- or 10 membered bicyclic heteroaryl,each of which is substituted with 0, 1, or 2 substituents independentlychosen from (a) and 0 or 1 substituents (b) where:

-   -   (a) is chosen from halogen, hydroxyl, amino, cyano, nitro,        —COOH, —CONH₂, ═NOH, C₁-C₄alkyl, C₁-C₄alkoxy, C₂-C₄alkanoyl,        C₁-C₄hydroxyalkyl, mono- and di-C₁-C₄alkylamino,        C₁-C₄alkylester, mono- or di-C₁-C₄alkylsulfonamide, mono- and        di-C₁-C₄alkylcarboxamide, C₁-C₂haloalkyl, C₁-C₂haloalkoxy, and    -   (b) is phenyl and 5- or 6-membered heteroaryl, each of which is        substituted with 0 or 1 or more substituents independently        chosen from halogen, hydroxyl, amino, —CHO, —COOH, —NH(C—O)H,        C₁-C₄alkyl, C₁-C₂alkoxy, mono- and di-(C₁-C₄alkyl)amino, mono-        and di-(C₁-C₄alkyl)carboxamide, C₁-C₄alkylester,        (C₁-C₄alkylester)amino, C₁-C₂haloalkyl, and C₁-C₂haloalkoxy.

(bb) The invention includes compounds and salts of (aa) in which (b) isphenyl, pyridyl, thiazolyl, pyrrolyl, or imidazolyl, each of which issubstituted with 0 or 1 or more substituents independently chosen fromhalogen, hydroxyl, amino, —CHO, —COON, —NH(C═O)H, C₁-C₄alkyl,C₁-C₂alkoxy, mono- and di-(C₁-C₄alkyl)amino, mono- anddi-(C₁-C₄alkyl)carboxamide, C₁-C₄alkylester, (C₁-C₄alkylester)amino,C₁-C₂haloalkyl, and C₁-C₂haloalkoxy.

(cc) R₈ and R₉ are taken together to form an optionally substituted 5-to 7-membered cycloalkyl ring.

(dd) R₈ and R₉ are taken together to form a cyclopentyl ring.

(ee) The invention includes compounds and salts of (cc) in which R₈ andR₉ are taken together to form a cyclopentyl ring. E.g. the inventionincludes compounds and salts of Formula VI.

(ff) In certain embodiments the invention includes compounds and saltsof Formula VI in which

-   -   R₃ is C₁-C₄alkyl, (C₃-C₇cycloalkyl)C₀-C₄alkyl, or        (phenyl)C₀-C₂alkyl, each of which is substituted with 0 to 3        substituents independently chosen from halogen, hydroxyl,        C₁-C₂alkyl, C₁-C₂alkoxy, mono- and di-C₁-C₂alkylamino,        C₁-C₂haloalkyl, and C₁-C₂haloalkoxy; and R₄ is hydrogen,        C₁-C₂alkyl, or C₁-C₂alkoxy; or    -   R₃ and R₄ are be joined to form a cyclopropyl ring, which is        substituted with 0 to 2 substituents independently chosen from        vinyl, C₁-C₂alkyl, and C₁-C₂alkoxy.    -   R₁₁ is C₁-C₆alkyl, (C₃-C₇cycloalkyl)C₀-C₄alkyl, or        (phenyl)C₀-C₂alkyl, each of which is substituted with 0 to 3        substituents independently chosen from halogen, hydroxyl,        C₁-C₂alkyl, C₁-C₂alkoxy, mono- and di-C₁-C₂alkylamino,        C₁-C₂haloalkyl, and C₁-C₂haloalkoxy; and R₁₂ is hydrogen,        C₁-C₂alkyl, or C₁-C₂alkoxy; or    -   R₁₁ and R₁₂ are be joined to form a cyclopropyl ring, which is        substituted with 0 to 2 substituents independently chosen from        vinyl, C₁-C₂alkyl, and C₁-C₂alkoxy; and    -   R₅, R₆, R₇, and R₁₀ are each independently hydrogen, methyl or        methoxy.

(gg) The invention also includes compounds and salts of Formula VII

(hh) The invention includes compounds and salts of Formula VIII

where D is a alkyl or alkenyl group having 6 to 10 carbon atoms;

-   R₄ and R₁₂ are independently hydrogen or methyl; and-   R₅, R₆, R₇, and R₁₀ are independently hydrogen or methyl.

(ii) The invention includes compounds and salts of Formula IX

where D is a alkyl or alkenyl group having 6 to 10 carbon atoms.

-   R₄ and R₁₂ are independently hydrogen or methyl; and-   R₅, R₆, R₇, and R₁₀ are independently hydrogen or methyl.    The R Variable

(jj) R is

(kk) R is a carboxamide group as shown in (jj) and R₁₃ is C₁-C₆alkyl orC₂-C₆alkenyl, each of which is substituted with 0 or 1 or moresubstituents independently chosen from halogen, hydroxyl, amino, cyano,amino, C₁-C₄alkoxy, mono or di-C₁-C₄alkylamino.

(ll) R is a carboxamide group as shown in (jj) and R₁₃ is(C₃-C₇cycloalkyl)C₀-C₂allyl, (aryl)C₀-C₂alkyl, (5- to 7-memberedheterocycloalkyl)C₀-C₂alkyl, or (heteroaryl)C₀-C₂alkyl, each of which issubstituted with 0 or 1 or more substituents independently chosen fromhalogen, hydroxyl, amino, cyano, amino, C₁-C₄alkyl, C₁-C₄alkoxy, mono ordi-C₁-C₄alkylamino, C₁-C₂haloalkyl, or C₁-C₂haloalkoxy.

(mm) R is a carboxamide group as shown in (jj) and R₁₃ isC₃-C₇cycloalkyl which is substituted with 0 or 1 or more substituentsindependently chosen from halogen, hydroxyl, amino, cyano, amino,C₁-C₂alkyl, C₁-C₂alkoxy, mono or di-C₁-C₂alkylamino.

(nn) R is a carboxamide group as shown in (jj) and R₁₃ is R₁₃ iscyclopropyl.

Any of the above definitions for the variables R and R₁-R₁₂ may becombined so long as a stable compound results. All such compounds areincluded in the scope of the invention.

Pharmaceutical Preparations

Compounds of the invention can be administered as the neat chemical, butare preferably administered as a pharmaceutical composition.Accordingly, the invention provides pharmaceutical formulationscomprising a compound or pharmaceutically acceptable salt of theinvention, together with at least one pharmaceutically acceptablecarrier.

Compounds of the invention may be administered orally, topically,parenterally, by inhalation or spray, sublingually, transdermally, viabuccal administration, rectally, as an ophthalmic solution, or by othermeans, in dosage unit formulations containing conventionalpharmaceutically acceptable carriers. The pharmaceutical composition maybe formulated as any pharmaceutically useful form, e.g., as an aerosol,a cream, a gel, a pill, a capsule, a tablet, a syrup, a transdermalpatch, or an ophthalmic solution. Some dosage forms, such as tablets andcapsules, are subdivided into suitably sized unit doses containingappropriate quantities of the active components, e.g., an effectiveamount to achieve the desired purpose.

Carriers include excipients and diluents and must be of sufficientlyhigh purity and sufficiently low toxicity to render them suitable foradministration to the patient being treated. The carrier can be inert orit can possess pharmaceutical benefits of its own. The amount of carrieremployed in conjunction with the compound is sufficient to provide apractical quantity of material for administration per unit dose of thecompound.

Classes of carriers include, but are not limited to binders, bufferingagents, coloring agents, diluents, disintegrants, emulsifiers,flavorants, glidents, lubricants, preservatives, stabilizers,surfactants, tableting agents, and wetting agents. Some carriers may belisted in more than one class, for example vegetable oil may be used asa lubricant in some formulations and a diluent in others. Exemplarypharmaceutically acceptable carriers include sugars, starches,celluloses, powdered tragacanth, malt, gelatin; talc, and vegetableoils. Optional active agents may be included in a pharmaceuticalcomposition, which do not substantially interfere with the activity ofthe compound of the present invention.

Binders are substances that bind or “glue” powders together and makethem cohesive by forming granules, thus serving as the “adhesive” in theformulation. Binders add cohesive strength to that already available inthe diluent or bulking agent. Examples of binders include starch,gelatin, natural sugars, corn sweeteners, natural and synthetic gumssuch as acacia, sodium alginate, carboxymethylcellulose, polyethyleneglycol and waxes. The amount of binder in the composition can range, forexample, from about 2 to about 20% by weight of the composition, or fromabout 3 to about 10% by weight, even more preferably from about 3 toabout 6% by weight.

Diluents include sugars such as lactose, sucrose, mannitol and sorbitol;starches derived from wheat, corn, rice and potato; and celluloses suchas microcrystalline cellulose. The amount of diluent in the compositionmay be, for example, about 10 to about 90% by weight of the totalcomposition, about 25 to about 75%, about 30 to about 60% by weight, orabout 12 to about 60%.

Disintegrants are materials added to a pharmaceutical composition tohelp it break apart (disintegrate) and release the active agent.Suitable disintegrants include starches; including “cold water soluble”modified starches such as sodium carboxymethyl starch; natural andsynthetic gums such as locust bean, karaya, guar, and tragacanth gum andagar; cellulose derivatives such as methylcellulose and sodiumcarboxymethylcellulose; microcrystalline celluloses and cross-linkedmicrocrystalline celluloses such as sodium croscarmellose; alginatessuch as alginic acid and sodium alginate; clays such as bentonites; andeffervescent mixtures. The amount of disintegrant in the composition canrange, for example, from about 2 to about 15% by weight of thecomposition or from about 4 to about 10% by weight.

Lubricants are substances added to a pharmaceutical formulation toenable the tablet, granules, etc. after it has been compressed, torelease from the; mold or die by reducing friction or wear. Examples oflubricants useful in pharmaceutical dosage fowls include boric acid,sodium benzoate, sodium acetate, sodium chloride, and the like.Lubricants are usually added at the very last step before tabletcompression, since they must be present on the surfaces of the granulesand in between them and the parts of the tablet press. The amount oflubricant in the composition can range, for example, from about 0.1 toabout 5% by weight of the composition, from about 0.5 to about 2%, orfrom about 0.3 to about 1.5% by weight. The amount of compound or saltof the invention in a unit dose may be generally varied or adjusted fromabout 1.0 milligram to about 1,000 milligrams, from about 1.0 to about900 milligrams, from about 1.0 to about 500 milligrams, or from about 1to about 250 milligrams, according to the particular application and thepotency of the compound. The actual dosage employed may be varieddepending upon the patient's age, sex, weight and severity of thecondition being treated.

Pharmaceutical compositions formulated for oral administration are oftenpreferred. These compositions contain between 0.1 and 99% of a compoundof the invention and usually at least about 5% (weight %) of a compoundof the invention. Some embodiments contain from about 25% to about 50%or from 5% to 75% of a compound of invention.

Liquids Formulations

Compounds of the invention can be incorporated into oral liquidpreparations such as aqueous or oily suspensions, solutions, emulsions,syrups, tinctures, syrups, or elixirs, for example. Moreover,formulations containing these compounds can be presented as a dryproduct, e.g. as granules or powders, for constitution with water orother suitable vehicle before use. Typical components of carriers forsyrups, elixirs, emulsions and suspensions include ethanol, glycerol,propylene glycol, polyethylene glycol, liquid sucrose, sorbitol andwater. Liquid preparations can contain conventional additives, such assuspending agents (e.g., sorbitol syrup, methyl cellulose,glucose/sugar, syrup, gelatin, hydroxyethyl cellulose, carboxymethylcellulose, aluminum stearate gel, and hydrogenated edible fats),emulsifying agents (e.g., lecithin, sorbitan monsoleate, or acacia),non-aqueous vehicles, which can include edible oils (e.g., almond oil,fractionated coconut oil, silyl esters, propylene glycol and ethylalcohol), and preservatives (e.g., methyl or propyl p-hydroxybenzoateand sorbic acid). Oral formulations may contain demulcent, flavoringagents, sweetening agents, such as sucrose or saccharin, taste-maskingagents, and coloring agents.

Suspensions

Aqueous suspensions contain the active material(s) in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example AVICEL RC-591, sodiumcarboxymethylcellulose, methylcellulose, hydropropylmethylcellulose,sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents, for example lecithin and polysorbate 80.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, n-propyl p-hydroxybenzoate, methyl parabens, propylparabens, and sodium benzoate.

Oily suspensions may be formulated by suspending the active ingredientsin a vegetable oil, for example peanut oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide palatable oralpreparations. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Emulsions

Pharmaceutical compositions of the invention may also be in the form ofoil-in-water emulsions. The oily phase may be a vegetable oil, forexample olive oil or peanut oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitol,anhydrides, for example sorbitan monoleate, and condensation products ofthe said partial esters with ethylene oxide, for example polyoxyethylenesorbitan monoleate.

Tablets and Capsules

Tablets typically comprise conventional pharmaceutically compatibleadjuvants as inert diluents, such as calcium carbonate, sodiumcarbonate, mannitol, lactose and cellulose; binders such as starch,gelatin and sucrose; disintegrants such as starch, alginic acid andcroscarmelose; lubricants such as magnesium stearate, stearic acid andtalc. Glidants such as silicon dioxide can be used to improve flowcharacteristics of the powder mixture. Coloring agents, such as the FD&Cdyes, can be added for appearance. Sweeteners and flavoring agents, suchas aspartame, saccharin, menthol, peppermint, and fruit flavors, areuseful adjuvants for chewable tablets. Capsules (including time releaseand sustained release formulations) typically comprise one or more soliddiluents disclosed above. The selection of carrier components oftendepends on secondary considerations like taste, cost, and shelfstability.

Such compositions may also be coated by conventional methods, typicallywith pH or time-dependent coatings, such that the subject compound isreleased in the gastrointestinal tract in the vicinity of the desiredtopical application, or at various times to extend the desired action.Such dosage forms typically include, but are not limited to, one or moreof cellulose acetate phthalate, polyvinylacetate phthalate,hydroxypropyl methylcellulose phthalate, ethyl cellulose, Eudragitcoatings, waxes and shellac.

Formulations for oral use may also be presented as hard or soft shellcapsules. A capsule is a dosage form administered in a special containeror enclosure containing an active agent. The active agent may be presentin solid, liquid, gel, or powder form, or any other pharmaceuticallyacceptable form. A capsule shell may be made of methyl cellulose,polyvinyl alcohols, or denatured gelatins or starch or other material.Hard shell capsules are typically made of blends of relatively high gelstrength bone and pork skin gelatins. Soft shell capsule shells areoften made of animal or plant gelatins. The capsule itself may containsmall amounts of dyes, opaquing agents, plasticizers and preservatives.

The active agent in a capsule may be mixed with an inert solid diluent,for example, calcium carbonate, calcium phosphate or kaolin, or in thecase of soft gelatin capsules the active ingredient may be mixed withwater or an oil medium, for example peanut oil, liquid paraffin or oliveoil.

Injectable and Parenteral Formulations

Pharmaceutical compositions may be in the form of a sterile injectableaqueous or oleaginous suspension. This suspension may be formulatedaccording to the known art using those suitable dispersing or wettingagents and suspending agents that have been mentioned above. The sterileinjectable preparation may also be sterile injectable solution orsuspension in a non-toxic parentally acceptable diluent or solvent, forexample as a solution in 1,3-butanediol. Among the acceptable vehiclesand solvents that may be employed are water, Ringer's solution, andisotonic sodium chloride solution. In addition, sterile, fixed oils areconventionally employed as a solvent or suspending medium. For thispurpose any bland fixed oil may be employed including synthetic mono- ordiglycerides. In addition, fatty acids such as oleic acid are useful inthe preparation of injectables.

Compounds of the invention may be administered parenterally in a sterilemedium. Parenteral administration includes subcutaneous injections,intravenous, intramuscular, intrathecal injection or infusiontechniques. The drug, depending on the vehicle and concentration used,can either be suspended or dissolved in the vehicle. Advantageously,adjuvants such as local anesthetics, preservatives and buffering agentscan be dissolved in the vehicle. In compositions for parenteraladministration the carrier typically comprises least about 90% by weightof the total composition.

Methods of Treatment

The invention includes methods of preventing and treating hepatitis Cinfections, by providing an effective amount of a compound of theinvention to patient at risk for hepatitis C infection or infected witha hepatitis C virus.

The pharmaceutical combinations disclosed herein are useful forpreventing and treating hepatitis C infections in patients. An effectiveamount of a pharmaceutical combination of the invention may be an amountsufficient to (a) prevent hepatitis C or a symptom of a hepatitis C fromoccurring in a patient who may be predisposed to hepatitis C but has notyet been diagnosed as having it or prevent diseases that may beassociated with or caused by a primary hepatitis C infection (such asliver fibrosis that can result in the context of chronic HCV infection);(b) inhibit the progression of hepatitis C; and (c) cause a regressionof the hepatitis C infection. An amount of a pharmaceutical compositioneffect to inhibit the progress or cause a regression of hepatitis Cincludes an amount effective to stop the worsening of symptoms ofhepatitis C or reduce the symptoms experienced by a patient infectedwith the hepatitis C virus. Alternatively a halt in progression orregression of hepatitis C may be indicated by any of several markers forthe disease. For example, a lack of increase or reduction in thehepatitis C viral load or a lack of increase or reduction in the numberof circulating HCV antibodies in a patient's blood are markers of a haltin progression or regression of hepatitis C infection. Other hepatitis Cdisease markers include aminotransferase levels, particularly levels ofthe liver enzymes AST and ALT. Normal levels of AST are from 5 to 40units per liter of serum (the liquid part of the blood) and normallevels of ALT are from 7 to 56 units per liter of serum. These levelswill typically be elevated in a HCV infected patient. Disease regressionis usually marked by the return of AST and ALT levels to the normalrange.

Symptoms of hepatitis C that may be affected by an effective amount of apharmaceutical combination of the invention include decreased liverfunction, fatigue, flu-like symptoms: fever, chills, muscle aches, jointpain, and headaches, nausea, aversion to certain foods, unexplainedweight loss, psychological disorders including depression, tenderness inthe abdomen, and jaundice.

“Liver function” refers to a normal function of the liver, including,but not limited to, a synthetic function including synthesis of proteinssuch as serum proteins (e.g., albumin, clotting factors, alkalinephosphatase, aminotransferases (e.g., alanine transaminase, aspartatetransaminase), 5′-nucleosidase, y glutaminyltranspeptidase, etc.),synthesis of bilirubin, synthesis of cholesterol, and synthesis of bileacids; a liver metabolic function, including carbohydrate metabolism,amino acid and ammonia metabolism, hormone metabolism, and lipidmetabolism; detoxification of exogenous drugs; and a hemodynamicfunction, including splanchnic and portal hemodynamics.

An effective amount of a combination described herein will also providea sufficient concentration of the active agents in the concentrationwhen administered to a patient. A sufficient concentration of an activeagent is a concentration of the agent in the patient's body necessary toprevent or combat the infection. Such an amount may be ascertainedexperimentally, for example by assaying blood concentration of theagent, or theoretically, by calculating bioavailability. The amount ofan active agent sufficient to inhibit viral infection in vitro may bedetermined with a conventional assay for viral infectivity such as areplicon based assay, which has been described in the literature.

The invention also includes using pharmaceutical combinations comprisinga compound of the invention and at least one additional active agent inprophylactic therapies. In the context of prophylactic or preventativetreatment an effective amount of a compound of the invention is anamount sufficient to significantly decrease the patient's risk ofcontracting a hepatitis C infection.

Methods of treatment include providing certain dosage amounts of acompound of the invention and the at least one additional active agentto a patient. Dosage levels of each active agent of from about 0.1 mg toabout 140 mg per kilogram of body weight per day are useful in thetreatment of the above-indicated conditions (about 0.5 mg to about 7 gper patient per day). The amount of active ingredient that may becombined with the carrier materials to produce a single dosage form willvary depending upon the patient treated and the particular mode ofadministration. Dosage emit forms will generally contain between fromabout 1 mg to about 500 mg of each active agent. In certain embodiments25 mg to 500 mg, or 25 mg to 200 mg of a compound of the invention areprovided daily to a patient. When the additional active agent is NM 283(valopicitabine), 100 mg to 1000 mg/day, or 200 mg to 800 mg/day, or 200to 400 mg/day of either of those agents are typically provided to thepatient. When the additional active agent is VX-950, 1000 mg to 3750mg/day, or 1200 mg to 1800 mg/day are administered to the patient.Treatment regiments in which VX-950 is an additional active agent andabout 350 to about 450 mg or about 700 to about 800 mg of VX-950 areadministered to a patient three times per day or about 350 to about 450mg or about 700 to about 800 mg is administered every 12 hours areparticularly included in the invention.

Frequency of dosage may also vary depending on the compound used and theparticular disease treated. However, for treatment of most infectiousdisorders, a dosage regimen of 4 times daily or less is preferred and adosage regimen of 1 or 2 times daily is particularly preferred.

It will be understood, however that the specific dose level for anyparticular patient will depend upon a variety of factors including theactivity of the specific compound employed, the age, body weight,general health, sex, diet, time of administration, route ofadministration, and rate of excretion, drug combination and the severityof the particular disease undergoing therapy.

Combination Methods

The invention includes methods of treatment in which a compound or saltof the invention is provided together with one or more additional activeagents. In certain embodiments the active agent (or agents) is an HCVprotease inhibitor or HCV polymerase inhibitor. For example the proteaseinhibitor may be telaprevir (VX-950) and the polymerase inhibitor may bevalopicitabine, or NM 107, the active agent which valopicitabine isconverted into in vivo.

According to the methods of the invention, the compound of the inventionand an additional active agent may be: (1) co-formulated andadministered or delivered simultaneously in a combined formulation; (2)delivered by alternation or in parallel as separate formulations; or (3)by any other combination therapy regimen known in the art. Whendelivered in alternation therapy, the methods of the invention maycomprise administering or delivering the compound of The invention andan additional active agent sequentially, e.g., in separate solution,emulsion, suspension, tablets, pills or capsules, or by differentinjections in separate syringes. In general, during alternation therapy,an effective dosage of each active ingredient is administeredsequentially, i.e., serially, whereas in simultaneous therapy, effectivedosages of two or more active ingredients are administered together.Various sequences of intermittent combination therapy may also be used.

In certain embodiments method of treatment includes providing a patientwith a compound of Formula I and an interferon such as a pegylatedinterferon or interferon gamma. The interferon may be the only compoundprovided with the compound of the invention or may be provided with anadditional active agent that is not an interferon.

The invention methods of treatment and pharmaceutical combinationsincluding compounds of the invention any one or combination of thefollowing compounds and substances as an additional active agent:

Caspase inhibitors: IDN 6556 (Idun Pharmaceuticals)

Cyclophilin Inhibitors: NIM811 (Novartis) and DEBIO-025 (Debiopharm)

Cytochrome P450 monooxygenase inhibitors: ritonavir (WO 94/14436),ketoconazole, troleandomycin, 4-methylpyrazole, cyclosporin,clomethiazole, cimetidine, itraconazole, fluconazole, miconazole,fluvoxamine, fluoxetine, nefazodone, sertraline, indinavir, nelfinavir,amprenavir, fosamprenavir, saquinavir, lopinavir, delavirdine,erythromycin, VX-944, and VX-497. Preferred CYP inhibitors includeritonavir, ketoconazole, troleandomycin, 4-methylpyrazole, cyclosporin,and clomethiazole

Glucocorticoids: hydrocortisone, cortisone, prednisone, prednisolone,methylprednisolone, triamcinolone, paramethasone, betamethasone, anddexamethasone

Hematopoietins: hematopoietin-1 and hematopoietin-2. Other members ofthe hematopoietin superfamily such as the various colony stimulatingfactors (e.g. (e.g. G-CSF, GM-CSF, M-CSF), Epo, and SCF (stem cellfactor)

Homeopathic Therapies: Milk Thistle, silymarin, ginseng, glycyrrhizin,licorice root, schisandra, vitamin C, vitamin E, beta carotene, andselenium

Immunomodulatory compounds: thalidomide, IL-2, hematopoietin, IMPDHinhibitors, for example Merimepodib (Vertex Pharmaceuticals Inc.),interferon, including natural interferon (such as OMNIFERON, Viragen andSUMIFERON, Sumitomo, a blend of natural interferons), natural interferonalpha (ALFERON, Hemispherx Biopharma, Inc.), interferon alpha nl fromlymphblastoid cells (WELLFERON, Glaxo Wellcome), oral alpha interferon,Peg-interferon, Peg-interferon alfa 2a (PEGASYS, Roche), recombinantinterferon alfa 2a (ROFERON, Roche), inhaled interferon alpha 2b (AERX,Aradigm), Peg-interferon alpha 2b (ALBUFERON, Human GenomeSciences/Novartis, PEGINTRON, Schering), recombinant interferon alfa 2b(INTRON A, Schering), pegylated interferon alfa 2b (PEG-INTRON,Schering, VIRAFERONPEG, Schering), interferon beta-1a (REBIF, Serono,Inc. and Pfizer), consensus interferon alpha (INFERGEN, ValeantPharmaceutical), interferon gamma-1b (ACTIMMUNE, Intermune, Inc.),un-pegylated interferon alpha, alpha interferon, and its analogs, andsynthetic thymosin alpha 1 (ZADAXIN, SciClone Pharmaceuticals Inc.)

Immunosupressants: sirolimus (RAPAMUNE, Wyeth)

Interleukins: (IL-1, IL-3, IL-4, IL-5, IL-6, IL-10, IL-11, IL-12), LIF,TGF-beta, TNF-alpha) and other low molecular weight factors (e.g.AcSDKP, pEEDCK, thymic hormones, and minicytokines)

Interferon Enhancers: EMZ702 (Transition Therapeutics)

IRES inhibitors: VGX-410C (VGX Pharma)

Monoclonal and Polyclonal antibodies: XTL-6865 (XTL), HuMax-HepC(Genmab), Hepatitis C Immune Globin (human) (CIVACIR, NabiBiopharmaceuticals)

Nucleoside analogues: Lamivudine (EPIVIR, 3TC, GlaxoSmithKline), MK-0608(Merck), zalcitabine (HIVID, Roche US Pharmaceuticals), ribavirin(including COPEGUS (Roche), REBETOL (Schering), VILONA (ICNPharmaceuticals, and VIRAZOLE (ICN Pharmaceuticals), and viramidine(Valeant Pharmaceuticals), an amidine prodrug of ribavirin. Combinationsof nucleoside analogues may also be employed.

Non-nucleoside inhibitors: PSI-6130 (Roche/Pharmasset), delaviridine(RESCRIPTOR, Pfizer), and HCV-796 (Viropharm)

P7 protein inhibitor: amantadine (SYMMETREL, Endo Pharmaceuticals, Inc.)

Polymerase inhibitors: NM283 (valopicitabine) (Idenix) and NM 107(Idenix).

Protease inhibitors: BILN-2061 (Boehringer Ingelheim), GW-433908(prodrug of Amprenavir, Glaxo/Vertex), indinavir (CRIXIVAN, Merck),ITMN-191 (Intermune/Array Biopharma), VX950 (Vertex) and combinationscomprising one or more of the foregoing protease inhibitors

RNA interference: SIRNA-034 RNAi (Sirna Therapeutics)

Therapeutic Vaccines: IC41 (Intercell), IMN-0101 (Imnogenetics), GI 5005(Globeimmune), Chronvac-C (Tripep/Inovio), ED-002 (Imnogenetics),Hepavaxx C (ViRex Medical)

TNF agonists: adalimumab (HUMIRA, Abbott), entanercept (ENBREL, Amgenand Wyeth), infliximab (REMICADE, Centocor, Inc.)

Tubulin inhibitors: Colchicine

Sphingosine-1-phosphate receptor modulators: FTY720 (Novartis)

TLR agonists: ANA-975 (Anadys Pharmaceuticals), TLR7 agonist (AnadysPharmaceuticals), CPG10101 (Coley), and TLR9 agonists including CPG 7909(Coley)

Cyclophilin Inhibitors: NIM811 (Novartis) and DEBIO-025 (Debiopharm)

Patients receiving hepatitis C medications are typically giveninterferon together with another active agent. Thus methods of treatmentand pharmaceutical combinations in which a compound of The invention isprovided together with an interferon, such as pegylated interferon alfa2a, as the additional active agents are included as embodiments.Similarly methods and pharmaceutical combinations in which ribavirin isan additional active agent are provided herein.

EXAMPLES

All nonaqueous reactions are performed under an atmosphere of dry argongas (99.99%). NMR spectra are recorded at ambient temperature using aBruker Avance 300 spectrometer (1H at 300.1 MHz and ¹³C at 75.5 MHz,).The chemical shifts for ¹H and ¹³C are reported in parts per million (δ)relative to external tetramethylsilane and are referenced to signals ofresidual protons in the deuterated solvent. Analytical HPLC is performedusing a Waters X-bridge C18 150×4.6 mm 3.5 μm column with a 20-minlinear gradient elution of increasing concentrations of acetonitrile inwater (5 to 95%) containing 0.1% trifluoroacetic acid with a flow rateof 1.0 mL/min and UV detection at 254 nm. Low-resolution mass spectraare recorded on a Thermo Finnigan Surveyor MSQ instrument (operating inAPCI mode) equipped with a Gilson liquid chromatograph. Unless notedotherwise, the quasi-molecular ions, [M+H]⁺, observed in thelow-resolution mass spectra are the base peaks.

This invention is further illustrated by the following examples thatshould not be construed as limiting.

Abbreviations

The following chemical abbreviations are used in Examples 1 to 3.Additional abbreviations used in these examples will be familiar tothose of skill in the art of organic chemical synthesis.

AcOEt Ethyl Acetate AIBN 2,2′-azobis(2-methylpropionitrile) BOCt-Butoxycarbonyl (BOC)₂O di-t-Butyl dicarbonate c. concentrated CDI1,1′-Carbonyldiimidazole DCM dichloromethane DIPEA DiisopropylethylamineEDCI 1-Ethyl-3-[3-dimethylaminopropyl]carbodiimide Et₃N triethylamineEtOH Ethanol Hex. Hexanes MeOH Methanol TFA Trifluoroacetic acid THFtetrahydrofuran TLC Thin Layer Chromatography

Example 1 Synthesis of Tertiary Amine Substituted Thiazoles (Compounds20-25)

Step 1. Preparation N1,N4-dicyclopropyl-2,3-dihydroxysuccinamide(Compound 2)

The mixture of cyclopropylamine (2.4 eq) and diethyl tartrate 1 isheated in a sealed tube at 70° C. overnight. The formed solid is washedwith diethyl ether to provide pure product 2 (>98% yield).

Step 2. Preparation N-cyclopropyl-2-hydroxy-3-nitrohexanamide (Compound4)

H₅IO₆ (5.8 g, 25.2 mmol) is added in several portions to a suspension of2 (5.7 g, 25 mmol) and silica gel (2.0 g) in 50 ml, methanol at 0° C.After completion of addition, the mixture is warmed to it for additional1.5 hr to complete the formation of compound 3. 1-Nitrobutane (6.2 g,1.2 eq) is added at 0° C. followed by Et₃N (7.4 mL). The mixture is keptstirring for 2.5 hr at 0° C. The solid is removed by filtration and thefiltrate concentrated. The resulting syrup is treated with water andethyl acetate and acidified with 1N HCl. The organic layer is collectedand the aqueous layer is extracted with ethyl acetate three times. Thecombined organic solvent is washed with brine and dried over magnesiumsulfate. The solvent is removed and the resulting solid was first washedwith hexane, then recrystallized with hexane-ether to give compound 4(>70% yield).

Step 3. Preparation of 3-amino-N-cyclopropyl-2-hydroxyhexanamide(Compound 5)

Compound 4 and Pd (10% on carbon, 30% weight of 4) in acetic acid ishydrogenated with H₂ (33 psi) overnight. The mixture is filtered throughCelite and the filtrate is concentrated. The remaining residue isevaporated with toluene twice and then diethyl ether is added to givecompound 5.

Step 4. Preparation or (1S,3aR,6aS)-ethyl-2-BOCsubstituted-4-oxooctahydrocyclopenta[c]pyrrole-1-carboxylate (Compound7)

Bu₃SnH (Aldrich, 36 ml, 136 mmol) followed by AIBN (2.5 g, 15 mmol) isadded into the tetra cyclic compound 6 (31.1 g, 100 mmol; prepared withMonn's method reported in JOC 59, 2773, 1994) dissolved in anhydroustoluene (160 ml). The mix is degassed and refluxed under Ar tillstarting material disappears 6 hr) as shown by TLC (Hex/AcOEt, 1/1).Toluene is evaporated under reduced pressure and the oil residue istreated with diethyl ether (100 ml) and HCl aq. solution (1.0 N, 110ml). After vigorous stirring for over night, diethyl ether is separated.The aqueous layer was extracted with diethyl ether (100 ml×6) and thenused for next step.

(BOC)₂O (32.7 g, 150 mmol) dissolved in THF is added into the aqueoussolution obtained above, (100 ml) followed by NaOH aq (4.5 g in water 10ml). The mixture is then cooled in an ice bath. The mix is stirred for 1hr and NaCl (s, 30 g) is then added and stirred. The organic layer andthe aqueous layer are extracted with AcOEt (50 ml×3). The combinedorganic layer is washed with brine and dried over anhydrous sodiumsulfate. After evaporation, the oily crude is applied to silica gelchromatography with Hex/AcOEt (4:1 to 3:1) as eluant. Compound 7 (25 g,84%) is obtained as colorless liquid.

Step 5. Preparation of (1S,3aR,6aS)-ethyl2-BOC-4-hydroxyoctahydrocyclopenta[c]pyrrole-1-carboxylate (Compound 8)

NaBH₄ (2.6 g, 67.3 mmol) is added in three portions at 0° C. withstirring to a solution of compound 7 (20 g, 67.3 mmol) in EtOH (150 ml).The mix is stirred for additional 1 hr for completion before AcOH (10ml) is carefully added at 0° C. to quench the reaction. EtOH is removedby evaporation and the residue is dissolved in AcOEt (300 ml) to performthe aqueous work up. After washing with sat NaHCO₃ (100 ml×2) and brine(100 ml) and drying over NaSO₄, the AcOEt solution is passed through ashort bed of silica gel. The solvent is evaporated to give compound 8(19.2 g, 95%) as colorless syrup.

Step 6. Preparation of (1S,3aR,6aS)-ethyl2-BOC-4-(phenoxycarbonothioyloxy)octahydrocyclopenta[c]pyrrole-1-carboxylate(Compound 9)

O-Phenyl chlorothionoformate (10 ml, 70.6 mmol) is added with stirringat 0° C. under Ar atmosphere, into the solution of compound 8 (19.2 g,64.2 mmol) in DCM (150 ml), followed by pyridine (6.22 ml, 77.04 mmol).The mix is stirred at rt over night. Methanol (5 ml) is added to destroyexcessive phenyl chlorothionocabonate before performing an aqueous workup. The organic layer is washed with 1N HCl (100 ml), water (100 ml),sat NaHCO₃ (100 ml), and brine (100 ml) in sequence. After drying overNaSO₄, the solvent is removed by evaporation to give compound 9 (27.5 g,99%) as golden color syrup.

Step 7. Preparation of (1S,3aR,6aS)-ethyl2-BOC-octahydrocyclopenta[c]pyrrole-1-carboxylate (Compound 10)

Bu₃SnH (25 ml, 94.8 mmol) followed by AIBN (1.55 g, 9.48 mmol) is addedat room temperature into the solution of compound 9 (27.5 g, 63.2 mmol)in benzene (60 ml). The mix is degassed twice and stirred at refluxtemperature under Ar for 4 hr. Volatiles are evaporated and the residueis applied to flash chromatography on silica gel with Hex/AcOEt(gradient 9/1 to 6/1) as eluant. Compound 10 (13 g, 73%) is obtained ascolorless liquid.

Step 8. Preparation of(1S,3aR,6aS)-2-BOC-octahydrocyclopenta[c]pyrrole-1-carboxylic acid(Compound 11)

Compound 10 (13 g, 46 mmol) is treated with 2N NaOH (44 ml) in EtOH (120ml) at rt for 3 hr. Volatiles are evaporated and the residue isdissolved in water (50 ml). 1N HCl is added to adjust pH to and theaqueous solution is back extracted with DCM (100 ml×3). The DCM isevaporated to give compound 11 (8.5 g, 72%) as colorless syrup.

Step 9. Preparation(1S,3aR,6aS)-N-(1-(cyclopropylamino)-2-hydroxy-1-oxohexan-3-yl)-2-BOC-octahydrocyclopenta[c]pyrrole-1-carboxamide(Compound 12)

Compound 11 (3.06 g, 12 mmol) is treated with EDCI (2.3 g, 12 mmol) andHOBt (1.84 g, 12 mmol) in DCM (50 ml) for 10 min. Compound 5 (1.79 g,9.6 mmol) followed by DIPEA (2.6 ml, 15 mmol) is added and the mix isstirred at rt over night. The mixture is evaporated to remove solventand the residue is treated with AcOEt (200 ml). After washing with 1NHCl 20 ml, sat. NaHCO₃ 50 ml×3, and brine (50 ml) in sequence and thendrying over NaSO₄, solvent is removed by evaporation to give compound 12(3.85 g, 95%) as a white foam.

Step 10. Preparation of(1S,3aR,6aS)—N-(1-(cyclopropylamino)-2-hydroxy-1-oxohexan-3-yl)octahydrocyclopenta[c]pyrrole-1-carboxamide(Compound 13)

Compound 12 (0.845 g, 2 mmol) is treated with TFA (5 ml) in DCM (10 ml)at 0° C. for 2 hr. The mixture is evaporated to dryness. The yellowsyrup so obtained is dissolved in DCM (20 ml) to make a stock solutionof 13 (0.1 M) for the next step.

Step 11. Preparation of(1S,3aR,6aS)—N-(1-(cyclopropylamino)-2-hydroxy-1-oxohexan-3-yl)-2-(2-(piperidin-1-yl)propanoyl)octahydrocyclopenta[c]pyrrole-1-carboxamideand related compounds (Compound 14-19)

2-Piperidin-1-yl-propionic acid hydrochloride (63 mg, 0.4 mmol) istreated with EDCI (77 mg, 0.4 mmol) and HOBt (61 mg, 0.4 mmol) in DCM (2ml) for 10 min. Stock solution of compound 13 (0.1 M, 4 ml) followed byDIPEA (0.122 ml, 0.7 mmol) is added and the mix is stirred at rt overnight. Aqueous work up with AcOEt extraction and sat. NaHCO₃ wash givescompound 14 (crude, 68 mg) after removal of solvent. Starting fromcorresponding acids, compounds 15-19 were prepared with similar method.

Step 12. Preparation of(1S,3aR,6aS)—N-(1-(cyclopropylamino)-1,2-dioxohexan-3-yl)-2-(2-(piperidin-1-yl)propanoyl)octahydrocyclopenta[c]pyrrole-1-carboxamideand related compounds (Compound 20-25)

Compound 14 (68 mg) is treated with Dess-Martin reagent (100 mg) in DCM(5 ml) at rt for 2 hr. The mix is concentrated and then applied to HPLCfor purification. Compound 20 (17 mg) is obtained as its TFA salt.Compounds 21-25 are obtained via similar methods.

Example 2 Synthesis of(3R,5S)-5-(1-(cyclopropylamino)-1,2-dioxohexan-3-ylcarbamoyl)-1-(2-(piperidin-1-yl)propanoyl)pyrrolidin-3-yl3,4-dihydroisoquinoline-2(1H)-carboxylate (Compound 33)

The syntheses of compounds 33 and 34 is depicted in Scheme 2.

Step 1. Preparation of (3R,5S)-5-(methoxycarbonyl)-1-BOC-pyrrolidin-3-yl3,4-dihydroisoquinoline-2(1H)-carboxylate (Compound 27)

Add Boc-trans-L-4-hydroxyproline methyl ester 26 (2.45 g, 10 mmol)dissolved in DCM (15 ml) dropwise into CDI (1.62 g, 10 mmol) solution inDCM (80 ml) at 0° C. After stirring at rt for 2 hr,1,2,3,4-tetrahydroisoquinoline (0.543 g, 4.08 mmol) in DCM (5 ml) isadded and the mixture is stirred over night. The solvent is removed andresidue applied to aqueous workup. The ethyl acetate (200 ml) layer iswashed with 1N HCl (20 ml×2), sat. sodium bicarbonate aqueous (30 ml×3),brine (10 ml) sequentially and dried over anhydrous sodium sulfate.Filter and remove the solvent by evaporation to provide compound 27(3.89 g, 96%) as yellow syrup.

Step 2. Preparation of(2S,4R)-1-BOC-4-(1,2,3,4-tetrahydroisoquinoline-2-carbonyloxy)pyrrolidine-2-carboxylicacid (Compound 28)

Compound 27 (3.89 g, 9.6 mmol) in MeOH (35 ml) is treated with 2N NaOH(15 ml) at rt for 3 hr. Solvent is removed by evaporation and theresidue dissolved in water (50 ml). Add c. HCl to adjust pH to ˜2. Theprecipitate is collected by filtration and dried to give compound 28(3.34 g, 89%) as a white powder.

Step 3. Preparation of(3R,5S)-5-(1-(cyclopropylamino)-2-hydroxy-1-oxohexan-3-ylcarbamoyl)-1-BOC-pyrrolidin-3-yl-3,4-dihydroisoquinoline-2(1H)-carboxylate(Compound 29)

Compound 28 (2.67 g, 6.85 mmol) is treated with EDCI (1.33 g, 6.85 mmol)and HOBt (1.06 g, 6.85 mmol) in DCM (50 ml) for 10 min. Compound 5 (1.15g, 6.2 mmol) followed by DIPEA (2.2 ml, 12.4 mmol) is added and the mixis stirred at rt over night. The mixture is evaporated to remove solventand the residue treated with AcOEt (100 ml). After washing with 1N HCl20 ml, sat. NaHCO₃ 30 ml×3, and brine 50 ml in sequence and then dryingover NaSO₄, solvent is removed by evaporation to give compound 29 (2.6g, 75%) as white foam.

Step 4. Preparation of(3R,5S)-5-(1-(cyclopropylamino)-2-hydroxy-1-oxohexan-3-ylcarbamoyl)pyrrolidin-3-yl-3,4-dihydroisoquinoline-2(1H)-carboxylate(Compound 30)

Compound 29 (2.6 g, 4.65 mmol) in DCM (25 ml) is treated with TFA (25ml) at 0° C. for 1 hr. Volatiles are evaporated to dryness. The residueis dissolved in AcOEt (100 ml) and washed with sat. NaHCO₃ (10 ml×3) andbrine (2 ml). After drying over NaSO₄, the solvent is removed byevaporation to give compound 30 (1.75 g, 82%) as pale yellow powder.

Step 5. Preparation of(3R,5S)-5-(1-(cyclopropylamino)-2-hydroxy-1-oxohexan-3-ylcarbamoyl)-1-(2-(piperidin-1-yl)propanoyl)pyrrolidin-3-yl-3,4-dihydroisoquinoline-2(1H)-carboxylateand related compound (Compound 31 and 32)

2-Piperidin-1-yl-propionic acid hydrochloride (58 mg, 0.3 mmol) istreated with EDCI (63 mg, 0.3 mmol) and HOBt (46 mg, 0.3 mmol) in DCM (5ml) for 10 min. Compound 30 (0.138 g, 0.3 mmol) followed by DIPEA (0.104ml, 0.6 mmol) is added and the mix is stirred at rt over night. Aqueouswork up with AcOEt extraction and sat NaHCO₃ wash provides compound 31(crude, 182 mg) after removal of solvent. Starting form correspondingacid, compound 32 (crude, 170 mg) is prepared with similar method.

Step 6. Preparation of(3R,5S)-5-(1-(cyclopropylamino)-1,2-dioxohexan-3-ylcarbamoyl)-1-(2-(piperidin-1-yl)propanoyl)pyrrolidin-3-yl-3,4-dihydroisoquinoline-2(1H)-carboxylate(Compound 33 and 34)

Compound 31 (182 mg) is treated with Dess-Martin reagent (170 mg) in DCM(5 ml) at rt for 2 hr. The mix is concentrated and then applied to HPLCfor purification. Compound 33 (47 mg) is obtained as pale yellow foam.Compounds 34 (36 mg) is obtained via a similar method.

Example 3 Synthesis of Macrocyclic Peptide Ketoamides

The synthesis of macro cyclic peptide keto amide is performed asdepicted in Scheme 3.

Example 4 Alternate Method for Macrocyclic Peptide Ketoamide Synthesis

The synthesis of macro cyclic peptide acid could be completed asdepicted in Scheme 4.

Example 5 Additional Tertiary Amine Substituted Peptides

The compounds shown in Table I (TABLE OF COMPOUNDS) are prepared by themethods given in Examples 1 to 4. Variations to these methods may benecessary to obtain certain compounds shown in Table I. Such variationsare routine in the art of synthetic organic chemistry and are readilyapparent to those of skill in the art. In Table I++ indicates inhibitionof HCV replicon replication of less than 100 micromolar, + indicatesinhibition of above 100 micromolar.

Cmp. # Structure Name EC50 35

N-(1-(cyclopropylamino)-1,2- dioxohexan-3-yl)-2-(2- (piperidin-1-yl)propanoyl)octahydrocyclopenta [c]pyrrole-1-carboxamide ++ 36

N-(1-(cyclopropylamino)-1,2- dioxohexan-3-yl)-2-(2-phenyl-2-(piperidin-1- yl)acetyl)octahydrocyclopenta[c] pyrrole-1-carboxamide ++37

2-(2-(azepan-1-yl)propanoyl)-N- (1-(cyclopropylamino)-1,2- dioxohexan-3-yl)octahydrocyclopenta[c]pyrrol e-1-carboxamide ++ 39

tert-butyl(2S)-1-(1-(1- (cyclopropylamino)-1,2-dioxohexan-3-ylcarbamoyl) hexahydrocyclopenta[c]pyrrol-2(1H)-yl)-3,3-dimethyl-1- oxobutan-2-ylcarbamate ++ 40

N-(1-(cyclopropylamino)-1,2- dioxohexan-3-yl)-2-(2- (pyrrolidin-1-yl)propanoyl)octahydrocyclopenta [c]pyrrole-1-carboxamide ++ 41

tert-butyl 4-(1-(1-(1- (cyclopropylamino)-1,2- dioxohexan-3-ylcarbamoyl)hexahydrocyclopenta [c]pyrrol-2(1H)-yl)-1-oxopropan-2-yl)piperazine-1- carboxylate ++ 42

N-(1-(cyclopropylamino)-1,2- dioxohexan-3-yl)-2-((R)-2- (piperidin-1-yl)propanoyl)octahydrocyclopenta [c]pyrrole-1-carboxamide + 43

N-(1-(cyclopropylamino)-1,2- dioxohexan-3-yl)-2-((R)-2- (pyrrolidin-1-yl)propanoyl)octahydrocyclopenta [c]pyrrole-1-carboxamide + 44

(3R,5S)-5-(1- (cyclopropylamino)-1,2- dioxohexan-3-ylcarbamoyl)-1-(2-(piperidin-1- yl)propanoyl)pyrrolidin-3-yl 3,4-dihydroisoquinoline-2(1H)- carboxylate ++ 45

(3R,5S)-S-(1- (cyclopropylamino)-1,2- dioxohexan-3-ylcarbamoyl)-1-(2-(pyrrolidin-l- yl)propanoyl)pyrrolidin-3-yl 3,4-dihydroisoquinoline-2(1H)- carboxylate + 46

N-(1-(cyclopropylamino)-1,2- dioxohexan-3-yl)-2-((S)-2- (piperidin-1-yl)propanoyl)octahydrocyclopenta [c]pyrrole-1-carboxamide ++ 47

N-(1-(cyclopropylamino)-1,2- dioxohexan-3-yl)-2-((S)-3,3-dimethyl-2-(piperidin-1- yl)butanoyl)octahydrocyclopenta[c]pyrrole-1-carboxamide ++ 48

N-(1-(cyclopropylamino)-1,2- dioxohexan-3-yl)-2-((S)-3,3-dimethyl-2-(piperidin-1- yl)butanoyl)octahydrocyclopenta[c]pyrrole-1-carboxamide ++ 49

N-(1-(cyclopropylamino)-1,2- dioxohexan-3-yl)-2-((S)-3-mcthyl-2-(piperidin-1 - yl)bulanoyl)octahydrocyclopenta[c]pyrrole-1-carboxamide ++ 50

ethyl 1-((2S,4R)-1-((S)-3,3- dimethyl-2-(piperidin-1-yl)butanoyl)-4-(7-methoxy-2- phenylquinolin-4- yloxy)pyrrolidine-2-carboxamido)-2- vinylcyclopropanecarboxylate ++ 51

ethyl 1-((2S,4R)-4-(7-methoxy- 2-phenylquinolin-4-yloxy)-1-((S)-2-phenyl-2-(piperidin-1- yl)acetyl)pyrrolidine-2- carboxamido)-2-vinylcyclopropanecarboxylate ++ 52

ethyl 1-((2S,4R)-4-(7-methoxy- 2-phenylquinolin-4-yloxy)-1-((S)-3-methyl-2-(piperidin-1- yl)butanoyl)pyrrolidine-2- carboxamido)-2-vinylcyclopropanecarboxylate ++ 53

ethyl 1-((2S,4R)-1-((S)-2- cyclohexyl-2-(piperidin-1-yl)acetyl)-4-(7-methoxy-2- phenylquinolin-4- yloxy)pyrrolidine-2-carboxamido)-2- vinylcyclopropanecarboxylate ++ 54

ethyl 1-((2S,4R)-4-(7-methoxy- 2-phenylquinolin-4-yloxy)-1-((S)-2-(piperidin-1- yl)propanoyl)pyrrolidine-2- carboxamido)-2-vinylcyclopropanecarboxylate ++ 55

1-((2S,4R)-1-((S)-3,3-dimethyl- 2-(piperidin-1-yl)butanoyl)-4-(7-methoxy-2-phenylquinolin-4- yloxy)pyrrolidine-2- carboxamido)-2-vinylcyclopropanecarboxylic acid ++ 56

1-((2S,4R)-4-(7-methoxy- 2-phenylquinolin-4-yloxy)-1-((S)-2-phenyl-2-(piperidin-1- yl)acetyl)pyrrolidine-2- carboxamido)-2-vinylcyclopropanecarboxylic acid + 57

1-((2S,4R)-4-(7-methoxy-2- phenylquinolin-4-yloxy)-1-((S)- 3-methyl-2-(piperidin-1- yl)butanoyl)pyrrolidine-2- carboxamido)-2-vinylcyclopropanecarboxylic acid ++ 58

1-((2S,4R)-1-((S)-2-cyclohexyl- 2-(piperidin-1-yl)acetyl)-4-(7-methoxy-2-phenylquinolin-4- yloxy)pyrrolidine-2- carboxamido)-2-vinylcyclopropanecarboxylic acid ++ 59

1-((2S,4R)-4-(7-methoxy-2- phenylquinolin-4-yloxy)-1-((S)-2-(piperidin-1- yl)propanoyl)pyrrolidine-2- carboxamido)-2-vinylcyclopropanecarboxylic acid + 60

(2R,6S,16aS,Z)-ethyl 2-(7- methoxy-2-phenylquinolin-4-yloxy)-5,16-dioxo-6-(piperidin- 1-yl)- 1,2,3,5,6,7,8,9,10,11,13a,14,14a,15,16,16a- hexadecahydrocyclopropa [e]pyrrolo[1,2-a][1,4]diazacyclopentadecine- 14a-carboxylatc ++ 61

(2R,6S,16aS,Z)-2-(7-methoxy-2- phenylquinolin-4-yloxy)-5,16-dioxo-6-(pipcridin-1-yl)- 1,2,3,5,6,7,8,9,10,11,13a,14,14a, 15,16,16a-hexadecahydrocyclopropa [e]pyrrolo[1,2-a] [1,4]diazacyclopentadecine-14a-carboxylic acid ++ 62

(2R,6S,13aR,14aS,16aS,Z)-ethyl 2-(7-methoxy-2-phenylquinolin-4-yloxy)-5,16-dioxo-6- (piperidin-1-yl)-1,2,3,5,6,7,8,9,10,11,13a,14,14a, 15,16,16a- hexadecahydrocyclopropa[e]pyrrolo[1,2-a] [1,4]diazacyclopentadecine- 14a-carboxylate ++ 63

(2R,6S,13aS,14aR,16aS,Z)-ethyl 2-(7-methoxy-2-phenylquinolin-4-yloxy)-5,16-dioxo-6- (piperidin-1-yl)-1,2,3,5,6,7,8,9,10,11,13a,14,14a, 15,16,16a- hexadecahydrocyclopropa[e]pyrrolo[1,2-a] [1,4]diazacyclopentadecine- 14a-carboxylate ++ 64

(2S,4R)-1-((S)-2-cyclohexyl-2- (piperidin-1-yl)acetyl)-N-(1-(cyclopropylsulfonylcarbamoyl)- 2-vinylcyclopropyl)-4-(7-methoxy-2-phenylquinolin-4- yloxy)pyrrolidine-2- carboxamide ++ 65

(2S,4R)-N-((1S,2R)-1- (cyclopropylsulfonylcarbamoyl)-2-vinylcyclopropyl)-4-(7- methoxy-2-phenylquinolin-4-yloxy)-1-((S)-2-phenyl-2- (piperidin-1- yl)acetyl)pyrrolidine-2-carboxamide ++ 66

(2S,4R)-N-((1R,2S)-1- (cyclopropylsulfonylcarbamoyl)-2-vinylcyclopropyl)-4-(7- methoxy-2-phenylquinolin-4-yloxy)-1-((S)-2-phenyl-2- (piperidin-1- yl)acetyl)pyrrolidine-2-carboxamide ++ 67

(2S,4R)-N-(1- (cyclopropylsulfonylcarbamoyl)- 2-vinylcyclopropyl)-4-(7-methoxy-2-phenylquinolin-4- yloxy)-1-((S)-2-(piperidin-1-yl)propanoyl)pyrrolidine-2- carboxamide ++ 68

(2S,4R)-N-(1- (cyclopropylsulfonylcarbamoyl)- 2-vinylcyclopropyl)-4-(7-methoxy-2-phenylquinolin-4- yloxy)-1-((S)-3-methyl-2- (piperidin-1-yl)butanoyl)pyrrolidine-2- carboxamide ++ 69

(2S,4R)-N-(1- (cyclopropylsulfonylcarbamoyl)- 2-vinylcyclopropyl)-4-(7-methoxy-2-phenylquinolin-4- yloxy)-1-((S)-4-methyl-2- (piperidin-1-yl)pentanoyl)pyrrolidine-2- carboxamide ++

Example 6 Assay for Identifying Compounds which Inhibit HCV Replication

Compounds claimed herein are tested for the ability to inhibit viralreplication of the Hepatitis C replicon in cultured cells in which theHCV replicon construct has been incorporated. The HCV replicon systemwas described by Bartenschlager, et. al (Science, 285, pp. 110-113(1999)). The replicon system is predictive of in vivo anti-HCV activity;compounds that are active in humans uniformly evidence activity in thereplicon assay.

In this assay HCV replicon containing cells are treated with differentconcentrations of the test compound to ascertain the ability of the testcompound to suppress replication of the HCV replicon. As a positivecontrol, HCV replicon-containing cells are treated with differentconcentrations of interferon alpha, a known inhibitor of HCVreplication. The replicon assay system includes NeomycinPhosphotransferase (NPT) as a component of the replicon itself in orderto detect the transcription of replicon gene products in the host cell.Cells in which the HCV replicon is actively replicating have high levelsof NPT; the level of NPT is proportional to HCV replication. Cells inwhich the HCV replicon is not replicating also have low levels of NPTand thus do not survive when treated with Neomycin. The NPT level ofeach sample is measured using a captured ELISA.

A protocol for testing compounds for the ability to inhibit viralreplication of the Hepatitis C replicon cultured cells in which thereplicon construct has been incorporated, follows.

6A. HCV Replicon and Replicon Expression

The HCV genome consists of a single ORF that encodes a 3000 amino acidpolyprotein. The ORF is flanked on the 5′ side by an untranslated regionthat serves as an internal ribosome entry site (IRES) and at the 3′ sideby a highly conserved sequence necessary for viral replication (3′-NTR).The structural proteins, necessary for viral infection, are located nearthe 5′ end of the ORF. The non-structural proteins, designated NS2 toNS5B comprise the remainder of the ORF.

The HCV replicon contains, 5′-3′, the HCV-IRES, the neomycinphosphotransferase (neo) gene, the IRES of encephalomyocarditis virus,which directs translation of HCV sequences NS3 to NS5B, and the 3′-NTR.The sequence of the HCV replicon has been deposited in GenBank(Accession no. AJ242652).

The replicon is transfected into Huh-7 cells using standard methods suchas electroporation.

6B. Cell Maintenance

The equipment and materials include, but are not limited to, Huh-7 HCVreplicon-containing cells, maintenance media (DMEM (Dulbecco's modifiedEagle media) supplemented with 10% FBS, L-glutamine, non-essential aminoacids, penicillin (100 units/ml), streptomycin (100 micrograms/ml), and500 micrograms/ml of Geneticin (G418), screening media (DMEMsupplemented with 10% FBS, L-glutamine, non-essential amino acids,penicillin (100 units/ml) and streptomycin (100 micrograms/ml)), 96 welltissue culture plates (flat bottom), 96 well plates (U bottom for drugdilution), Interferon alpha for positive control, fixation reagent (suchas methanol: acetone), primary antibody (rabbit anti-NPTII), secondaryantibody: Eu-N1 1, and enhancement solution.

HCV replicon-containing cells support high levels of viral RNA repliconreplication when their density is suitable. Over-confluency causesdecreased viral RNA replication. Therefore, cells must be kept growingin log phase in the presence of 500 micrograms/ml of G418. Generally,cells should be passed twice a week at 1: 4-6 dilution. Cell maintenanceis conducted as follows:

HCV replicon-containing cells are examined under a microscope to ensurethat cells growing well. Cells are rinsed once with PBS and 2 ml trypsinis added. The cell/trypsin mixture is incubated at 37° C. in a CO₂incubator for 3-5 minutes. After incubation 10 ml of complete media isadded to stop the trypsinization reaction. Cells are blown gently, putinto a 15 ml tube, and spun at 1200 rpm for 4 minutes. Thetrypsin/medium solution is removed. Medium (5 ml) is added and the cellsare mixed carefully. The cells are counted.

The cells are then seeded onto 96-well plates at a density of 6000-7500cells/100 microliters/well (6-7.5×10⁵ cells/10 nil/plate). The platesare then incubated at 37° C. in a 5% CO₂ incubator.

Cells are examined under a microscope approximated 24 hours afterseeding and prior to adding drugs. If counting and dilution wereperformed correctly, cells are 60-70% confluent and nearly all cellsshould attach and spread evenly in the well.

6C. Treatment of HCV-Replicon Containing Cells with Test Compound

HCV replicon-containing cells are rinsed with once PBS once; 2 mls oftrypsin are then added. Cells are incubated at 37° C. in a 5% CO₂incubator for 3-5 minutes. 10 mls of complete medium is added to stopthe reaction. Cells are blown gently, put into a 15 ml tube, and spun at1200 rpm for four minutes. The trypsin/medium solution is removed and 5mls of medium (500 ml DMEM (high glucose)) from BRL catalog #12430-054;50 mls 10% FBS, 5% Geneticin G418 (50 mg/ml, BRL catalog #10131-035), 5ml MEM non-essential amino acids (100×BRL #11140-050) and 5 ml pen-strep(BRL #15140-148) is added. The cells and media are mixed carefully

Cells are plated with screening medium (500 ml DMEM (BRL #21063-029), 50ml FBS (BRL #10082-147) and 5 ml MEM non-essential amino acid (BRL#11140-050) at 6000-7500 cells/100 μl/well of 96 well plate (6-7.5×10⁵cells/10 ml/plate). Plates are placed into 37° C. 5% CO₂ incubatorovernight. 6D. Assay

The following morning, drugs (test compounds or interferon alpha) arediluted in 96 well U bottom plates with media or DMSO/media, dependingon the final concentration chosen for screening. Generally for 6concentrations of each test compounds ranging from 10 micromolar to 0.03micromolar are applied. 100 μl of the test compound dilution is placedin wells of the 96 well plate containing the HCV replicon cells. Mediawithout drug is added to some wells as a negative controls. DMSO isknown to affect cell growth. Therefore, if drugs diluted in DMSO areused, all wells, including negative control (media only) and positivecontrol (interferon alpha) wells, must contain the same concentration ofDMSO, for single dose screening. The plates are incubated at 37° C. in ahumidified 5% CO₂ environment for three days.

On day four, the NTPII assay is quantitated. The medium is poured fromthe plates and the plates are washed once in 200 μl of PBS. The PBS isthen decanted and the plates tapped in a paper towel to remove anyremaining PBS. Cells are fixed in situ with 100 id/well of pre-cooled(−20° C.) methanol:acetone (1:1) and the plates are placed at −20° C.for 30 minutes.

The fixing solution is poured from the plates and the plates allowed toair-dry completely (approximately one hour). The appearance of the driedcell layer is recorded and the density of the cells in the toxic wellsis scored with the naked eye. Alternatively cell viability may beassessed using the MTS assay described below.

The wells are blocked with 200 μl of blocking solution (10% FBS; 3% NGSin PBS) for 30 minutes at room temperature. The blocking solution isremoved and 100 μl of rabbit anti-NPTII diluted 1:1000 in blockingsolution is added to each well. The plates are then incubated 45-60minutes at room temperature. After incubation, wells are washed sixtimes with PBS-0.05% Tween-20 solution. 100 μl of 1:15,000 dilutedEuropium (EU)-conjugated goat anti-rabbit in blocking buffer is added toeach well and incubated at room temperature for 30-45 minutes. Theplates are washed again and 100 μl of enhancement solution (Perkin Elmer#4001-0010) is added to each well. Each plate is shaken (approx. 30 rpm)in a plate shaker for three minutes. 95 μl is transferred from each wellto a black plate; the EU signal is quantitated in a Perkin-Elmer VICTORplate reader (EU-Lance).

Test Results: Compounds described in the “TABLE OF COMPOUNDS” is Example5 have been tested in an HCV replication assay, essentially as describedin this example.

Example 7 Cytotoxicity Assays

To insure that the decrease in replicon replication is due to compoundactivity against the HCV replicon rather than nonspecific toxicityassays are used to quantitate compound cytotoxicity.

Example 7A Cellular Protein Albumin Assay for Cytotoxicity

Cellular protein albumin measurements provide one marker ofcytotoxicity. The protein levels obtained from cellular albumin assaysmay also be used to provide a normalization reference for antiviralactivity of compounds. In the protein albumin assay HCVreplicon-containing cells are treated for three days with differentconcentrations of helioxanthin; a compound that is known to be cytotoxicat high concentrations. The cells are lysed and the cell lysate used tobind plate-bound goat anti-albumin antibody at room temperature (25° C.to 28° C.) for 3 hours. The plate is then washed 6 times with 1×PBS.After washing away the unbound proteins, mouse monoclonal anti-humanserum albumin is applied to bind the albumin on the plate. The complexis then detected using phosphatase-labeled anti-mouse IgG as a secondantibody.

Example 7B MTS Assay for Cytotoxicity

Cell viability may also be determined by CELLTITER 96 AQUEOUS ONESolution Cell Proliferation Assay (Promega, Madison Wis.), acolorimetric assay for determining the number of viable cells. In thismethod, before fixing the cells, 10-20 μl MTS reagent is added to eachwell according to manufacturer's instructions, plates are incubated at37° C. and read at OD 490 nm. During the incubation period living cellscovert the MTS reagent to a formazan product which absorbs at 490 nm.Thus the 490 nm absorbance is directly proportional to the number ofliving cells in culture.

A direct comparison of the Cellular Album and MTS methods fordetermining cytotoxicity may be obtained as follows: Cells are treatedwith different concentrations of test compound or Helioxanthin for athree day-period. Prior to lysis for detection album as described above,the MTS reagent is added according to manufacturer's instruction to eachwell and incubate at 37° C. and read at OD 490 nm. The cellular albumquantitation is then performed as described above.

What is claimed is:
 1. A compound or pharmaceutically acceptable salt ofthe formula

wherein R is hydroxyl, C₁-C₄alkoxy,

R₁ is, C₂-C₆alkenyl, C₃-C₇cycloalkyl, phenyl, heterocycloalkyl, or 5- or6-membered heteroaryl, each of which is optionally substituted; R₂ isC₁-C₆ alkyl or C₂-C₆alkenyl; each of which is optionally substituted; orR₁ and R₂ are joined to form a 5- to 7-membered heteretycloalkyl ring,which ring is substituted with 0 or 1 or more substituents independentlychosen from halogen, hydroxyl, C₁-C₄alkyl, C₁-C₄alkoxy, C₁-C₂haloalkyl,and C₁-C₂haloalkoxy; R₃ and R₁₁ are independently C₁-C₆alkyl,C₂-C₆alkenyl, C₂-C₆alkanoyl, or mono- or di- C₁-C₆alkylamino,(C₃-C₇cycloalkyl)C₀-C₄alkyl, (C₃-C₇cycloalkenyl)C₀-C₄alkyl,(phenyl)C₀-C₂alkyl, or (heterocycloalkyl)C₀-C₄alkyl, each of which issubstituted with 0 to 3 substituents independently chosen from halogen,hydroxyl, amino, cyano, —CONH₂, —COOH, C₁-C₄alkyl, C₂-C₄alkanoyl,C₁-C₄alkoxy, C₁-C₄alkylthio, mono- and di-C₁-C₄alkylamino,C₁-C₂haloalkyl, and C₁-C₂haloalkoxy, R₄ and R₁₂ are independentlyhydrogen, halogen, hydroxyl, amino, C₁-C₆alkyl, C₂-C₆alkenyl, orC₁-C₆alkoxy; or R₃ and R₄ are joined to form a 3- to 7-memberedcycloalkyl ring or 5- to 7-membered heterocycloalkyl ring, each of whichis substituted with 0 to 2 substituents independently chosen fromhalogen, hydroxyl, amino, cyano, vinyl, C₁-C₂alkyl, C₁-C₂alkoxy,trifluoromethyl, and trifluoromethoxy, or R₁₁ and R₁₂ are joined to forma 3- to 7-membered cycloalkyl ring or 5- to 7-membered heterocycloalkylring, each of which is substituted with 0 to 2 substituentsindependently chosen from halogen, hydroxyl, amino, cyano, vinyl,C₁-C₂alkyl, C₁-C₂alkoxy, trifluoromethyl, and trifluoromethoxy; or R₃ isa C₇-C₁₁ saturated or unsaturated hydrocarbon chain that is (i)covalently bound to R₁₁, where R₁₁ is a methylene group or (ii)covalently bound to a cycloalkyl group formed by R₁₁ and R₁₂ beingjoined to form a 3- to 7-membered cycloalkyl ring; R₅ R₆, R₇, and R₁₀are independently hydrogen, halogen, cyano, amino, hydroxyl, C₁-C₂alkyl,C₁-C₂alkoxy, C₁-C₂haloalkyl, or C₁-C₂haloalkoxy; R₈ is a group of theformula —(CH₂)_(n)Y—Z, where n is 0, 1, or 2, and R₉ is hydrogen,halogen, amino, C₁-C₂alkyl, or C₁-C₂alkoxy; or R₈ and R₉ are takentogether to form an optionally substituted 5- to 7-membered cycloalkylring; R₁₃ is C₁-C₆alkyl, C₂-C₆alkenyl, each of which is substituted with0 or 1 or more substituents independently chosen from halogen, hydroxyl,amino, cyano, amino, C₁-C₄alkoxy, mono or di-C₁-C₄alkylamino or R₁₃ is(C₃-C₇cycloalkyl)C₀-C₂alkyl, (aryl)C₀-C₂alkyl, (5- to 7-memberedheterocycloalkyl)C₀-C₂alkyl, or (heteroaryl)C₀-C₂alkyl, each of which issubstituted with 0 or 1 or more substituents independently chosen fromhalogen, hydroxyl, amino, cyano, amino, C₁-C₄alkyl, C₁-C₄alkoxy, mono ordi-C₁-C₄alkylamino, C₁-C₂haloalkyl, or C₁-C₂haloalkoxy; Y is absent,CR₁₄R₁₅, NR₁₆, S, —O—, —O(C═O)(NR₁₆)—, —OC₁₄R₁₅—, NH(C═O)(NR₁₆)—,—NR₁₆(C═O)CR₁₄R₁₅—, NH(S═O)(NR₁₆)—, or —O(C═O)—; where R₁₄ and R₁₅ areindependently hydrogen, hydroxyl, halogen, C₁-C₂alkyl, C₁-C₂alkoxy,C₁-C₂haloalkyl, or C₁-C₂haloalkoxy; and R₁₆ is hydrogen, C₁-C₂alkyl,C₁-C₂haloalkyl, or C₁-C₂haloalkoxy; and Z is C₁-C₆alkyl, C₂-C₆alkenyl,(mono- or bicyclic aryl)C₀-C₂alkyl, (mono- or bicyclicheteroaryl)C₀-C₂alkyl, (C₃-C₇cycloalkyl)C₀-C₂alkyl, (5- or 6-memberedheterocycloalkyl)C₀-C₂alkyl, partially unsaturated bicyclic heterocycle,tricyclic aryl, or tricyclic hetero aryl; each of which Z is substitutedwith 0 or 1 or more substituents independently chosen from halogen,hydroxyl, amino, cyano, —CONH₂, —COOH, mono- anddi-C₁-C₄alkylsulfonamide, mono- and di- C₁-C₄alkylearboxamide,C₁-C₄alkyl, C₂-C₄alkanoyl, C₁-C₄alkoxy, C₁-C₄alkylthio, mono- anddi-C₁-C₄alkylamino, C₁-C₄alkylester, C₁-C₂haloalkyl, andC₁-C₂haloalkoxy, and 0 or 1 (C₃-C₇cycloalkyl)C₀-C₂alkyl,(aryl)C₀-C₂alkyl, (phenyl)C₀-C₂alkoxy, (5- or 6-memberedheteroaryl)C₀-C₂alkyl, (5- or 6-membered heteroaryl)C₀-C₂alkoxy,indanyl, (5- or 6-membered heterocycloalkyl)C₀-C₂alkyl, or 9- or 10membered bicyclic heteroaryl, each of which is substituted with 0, 1, or2 substituents independently chosen from (a) and 0 or 1 substituents (b)where: (a) is chosen from halogen, hydroxyl, amino, cyano, nitro, —COOH,—CONH₂, ═NOH, C₁-C₄alkyl, C₁-C₄alkoxy, C₂-C₄alkanoyl, C₁-C₄hydroxyalkyl,mono- and di-C₁-C₄alkylamino, C₁-C₄alkylester, mono- or di-C₁-C₄alkylsulfonamide, mono- and di-C₁-C₄alkylearboxamide,C₁-C₂haloalkyl, C₁-C₂haloalkoxy, and (b) is phenyl and 5- or 6-memberedheteroaryl, each of which is substituted with 0 or 1 or moresubstituents independently chosen from halogen, hydroxyl, amino, —CHO,—COOH, —NH(C═O)H, C₁-C₄alkyl, C₁-C₂alkoxy, mono- anddi-(C₁-C₄alkyl)amino, mono- and di-(C₁-C₄alkyl)carboxamide,C₁-C₄alkylester, (C₁-C₄alkylester)amino, C₁-C₂haloalkyl, andC₁-C₂haloalkoxy.
 2. A compound or salt of claim 1, in which R₁ and R₂are joined to form a pyrrolidinyl, piperidinyl, or piperazinyl ring,each of which is substituted with 0, 1, or 2 substituents independentlychosen from halogen, hydroxyl, C₁-C₂alkyl, C₁-C₂alkoxy, trifluoromethyl,and trifluoromethoxy.
 3. A compound or salt of claim 1, in which: R₃ isC₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkanoyl, or mono- or di-C₁-C₆alkylamino, (C₃-C₇cycloalkyl)C₀-C₄alkyl,(C₃-C₇cycloalkenyl)C₀-C₄alkyl, (phenyl)C₀-C₂alkyl, or(heterocycloalkyl)C₀-C₄alkyl, each of which is substituted with 0 to 3substituents independently chosen from halogen, hydroxyl, amino, cyano,—CONH₂, —COOH, C₁-C₄alkyl, C₂-C₄alkanoyl, C₁-C₄alkoxy, C₁-C₄alkylthio,mono- and di-C₁-C₄alkylamino, C₁-C₂haloalkyl, and C₁-C₂haloalkoxy, andR₄ is hydrogen, C₁-C₆alkyl, C₂-C₆alkenyl, or C₁-C₆alkoxy.
 4. A compoundor salt of claim 1 in which R₃ and R₄ are joined to form a cyclopropylring, which is substituted with 0 to 2 substituents independently chosenfrom vinyl, C₁-C₂alkyl, and C₁-C₂alkoxy.
 5. A compound or salt of claim1, in which: R₁₁ is C₁-C₆alkyl, C₂-C₆alkenyl, C₂-C₆alkanoyl, or mono- ordi- C₁-C₆alkylamino, (C₃-C₇cycloalkyl)C₀-C₄alkyl,(C₃-C₇cycloalkenyl)C₀-C₄alkyl, (phenyl)C₀-C₂alkyl, or(heterocycloalkyl)C₀-C₄alkyl, each of which is substituted with 0 to 3substituents independently chosen from halogen, hydroxyl, amino, cyano,—CONH₂, —COOH, C₁-C₄alkyl, C₂-C₄alkanoyl, C₁-C₄alkoxy, C₁-C₄alkylthio,mono- and di-C₁-C₄alkylamino, C₁-C₂haloalkyl, and C₁-C₂haloalkoxy, andR₁₂ is hydrogen, C₁-C₆alkyl, C₂-C₆alkenyl, or C₁-C₆alkoxy.
 6. A compoundor salt of claim 1 in which R₃ and R₄ are joined to form a cyclopropylring, which is substituted with 0 to 2 substituents independently chosenfrom vinyl, C₁-C₂alkyl, and C₁-C₂alkoxy.
 7. A compound or salt of claim1 of the formula

where D is an alkyl or alkenyl group having 6 to 10 carbon atoms.
 8. Acompound or salt of claim 7 of the formula


9. A compound or salt of claim 1 of the formula

where D is an alkyl or alkenyl group having 6 to 10 carbon atoms.
 10. Acompound or salt of claim 9 of the formula


11. A compound or salt of claim 1, in which R₅ R₆, R₇, and R₁₀ are allhydrogen.
 12. A compound or salt of claim 1, in which R₈ is a group ofthe formula —(CH₂)_(n)Y—Z, where n is 0, 1, or 2, and R₉ is hydrogen,halogen, amino, C₁-C₂alkyl, or C₁-C₂alkoxy.
 13. A compound or salt ofclaim 12 in which n is 0 and Y is —O— or —O(C═O)—.
 14. A compound orsalt of claim 1, wherein Z is a group of the formula

wherein X₁, X₂, X₃, and X₄, are independently N or CH and no more thantwo of X₁-X₄ are N; R₂₁ represents from 0 to 3 groups independentlychosen from halogen, hydroxyl, amino, cyano, —CONH₂, —COOH, C₁-C₄alkyl,C₂-C₄alkanoyl, C₁-C₄alkoxy, C₁-C₄alkylthio, mono- anddi-C₁-C₄alkylamino, C₁-C₂haloalkyl, and C₁-C₂haloalkoxy. R₂₂ ishydrogen, halogen, hydroxyl, amino, cyano, —CONH₂, —COOH, C₁-C₄alkyl,C₂-C₄alkanoyl, C₁-C₄alkoxy, C₁-C₄alkylthio, mono- anddi-C₁-C₄alkylamino, C₁-C₄alkylester, C₁-C₂haloalkyl, andC₁-C₂haloalkoxy, or R₂₂ is (C₃-C₇cycloalkyl)C₀C₂alkyl,(phenyl)C₀C₂alkyl, (phenyl)C₀C₂alkoxy, (5- or 6-memberedheteroaryl)C₀-C₂alkyl, (5- or 6-membered heteroaryl)C₀-C₂alkoxy,indanyl, (5- or 6-membered heterocycloalkyl)C₀C₂alkyl, or 9- or 10membered bicyclic heteroaryl, each of which is substituted with 0, 1, or2 substituents independently chosen from (a) and 0 or 1 substituents (b)where (a) is chosen from halogen, hydroxyl, amino, cyano, nitro, —COOH,—CONH₂, CH₃(C═O)NH—, C₁-C₄alkyl, C₁-C₄alkoxy, C₁-C₄hydroxyalkyl, mono-and di-C₁-C₄alkylamino, C₁-C₄alkylester, mono- and di-C₁-C₄alkylsulfonamide, mono- and di-C₁-C₄alkylcarboxamide,C₁-C₂haloalkyl, and C₁-C₂haloalkoxy, and (b) is phenyl or 5- or6-membered heteroaryl, each of which is substituted with 0 or 1 or moreof halogen, hydroxyl, C₁-C₄alkyl, and C₁-C₂alkoxy.
 15. A compound orsalt of claim 1 of the formula


16. A compound or salt of claim 15 wherein R₃ is C₁-C₆alkyl,(C₃-C₇cycloalkyl)C₀-C₄alkyl, or (phenyl)C₀-C₂alkyl, each of which issubstituted with 0 to 3 substituents independently chosen from halogen,hydroxyl, C ₁-C₂alkyl, C₁-C₂alkoxy, mono- and di-C₁-C₂alkylamino,C₁-C₂haloalkyl, and C₁-C₂haloalkoxy; and R₄ is hydrogen, C₁-C₂alkyl, orC₁-C₂alkoxy; or R₃ and R₄ are joined to form a cyclopropyl ring, whichis substituted with 0 to 2 substituents independently chosen from vinyl,C₁-C₂alkyl, and C₁-C₂alkoxy; R₁₁ is C₁-C₆alkyl,(C₃-C₇cycloalkyl)C₀-C₄alkyl, or (phenyl)C₀-C₂alkyl, each of which issubstituted with 0 to 3 substituents independently chosen from halogen,hydroxyl, C ₁-C₂alkyl, C₁-C₂alkoxy, mono- and di-C₁-C₂alkylamino,C₁-C₂haloalkyl, and C₁-C₂haloalkoxy; and R₁₂ is hydrogen, C₁-C₂alkyl, orC₁-C₂alkoxy; or R₁₁ and R₁₂ are joined to form a cyclopropyl ring, whichis substituted with 0 to 2 substituents independently chosen from vinyl,C₁-C₂alkyl, and C₁-C₂alkoxy; and R₅, R₆, R₇, and R₁₀ are eachindependently hydrogen, methyl or methoxy.
 17. A compound or salt ofclaim 15 of the formula


18. A compound or salt of claim 15 of the formula

where D is an alkyl or alkenyl group having 6 to 10 carbon atoms; R₄ andR₁₂ are independently hydrogen or methyl; and R₅, R₆, R₇, and R₁₀ areindependently hydrogen or methyl.
 19. A compound or salt of claim 15 ofthe formula

where D is an alkyl or alkenyl group having 6 to 10 carbon atoms; R₄ andR₁₂ are independently hydrogen or methyl; and R₅, R₆, R₇, and R₁₀ areindependently hydrogen or methyl.
 20. A pharmaceutical compositioncomprising a compound or salt of claim 1, containing at least onepharmaceutically acceptable carrier.
 21. A method for treating hepatitisC infection in a subject having hepatitis C infection comprisingproviding an effective amount of a compound or salt of claim 1 to apatient in need of such treatment.
 22. A compound or salt of claim 1,wherein the compound is

N-(1-(cyclopropylamino)-1,2-dioxohexan-3-yl)-2-(2-(piperidin-1-yl)propanoyl)octahydrocyclopenta[c]pyrrole-1-carboxamide;N-(1-(cyclopropylamino)-1,2-dioxohexan-3-yl)-2-(2-phenyl-2-(piperidin-1-yl)acetyl)octahydrocyclopenta[c]pyrrole-1-carboxamide;2-(2-(azepan-1-yl)propanoyl)-N-(1-(cyclopropylamino)-1,2-dioxohexan-3-yl)octahydrocyclopenta[c]pyrrole-1-carboxamide; tert-butyl(2S)-1-(1-(1-(cyclopropylamino)-1,2-dioxohexan-3-ylcarbamoyl)hexahydrocyclopenta[c]pyrrol-2(1H)-yl)-3,3-dimethyl-1-oxobutan-2-ylcarbamate;N-(1-(cyclopropylamino)-1,2-dioxohexan-3-yl)-2-(2-(pyrrolidin-1-yl)propanoyl)octahydrocyclopenta[c]pyrrole-1-carboxamide; tert-butyl4-(1-(1-(1-(cyclopropylamino)-1,2-dioxohexan-3-ylcarbamoyl)hexahydrocyclopenta[c]pyrrol-2(1H)-yl)-1-oxopropan-2-yl)piperazine-1-carboxylate;N-(1-(cyclopropylamino)-1,2-dioxohexan-3-yl)-2((R)-2-(piperidin-1-yl)propanoyl)octahydrocyclopenta[c]pyrrole-1-carboxamide;N-(1-(cyclopropylamino)-1,2-dioxohexan-3-yl)-2((R)-2-(pyrrolidin-1-yl)propanoyl)octahydrocyclopenta[c]pyrrole-1-carboxamide;(3R,5S)-5-(1-(cyclopropylamino)-1,2-dioxohexan-3-ylcarbamoyl)-1-(2-(piperidin-1-yl)propanoyl)pyrrolidin-3-yl3,4-dihydroisoquinoline-2(1H)-carboxylate;(3R,5S)-5-(1-(cyclopropylamino)-1,2-dioxohexan-3-ylcarbamoyl)-1-(2-(pyrrolidin-1-yl)propanoyl)pyrrolidin-3-yl3,4-dihydroisoquinoline-2(1H)-carboxylate;N-(1-(cyclopropylamino)-1,2-dioxohexan-3-yl)-2((S)-2-(piperidin-1-yl)propanoyl)octahydrocyclopenta[c]pyrrole-1-carboxamide;N-(1-(cyclopropylamino)-1,2-dioxohexan-3-yl)-2((S)-3,3-dimethyl-2-(piperidin-1-yl)butanoyl)octahydrocyclopenta[c]pyrrole-1-carboxamide;N-(1-(cyclopropylamino)-1,2-dioxohexan-3-yl)-2((S)-3,3-dimethyl-2-(piperidin-1-yl)butanoyl)octahydrocyclopenta[c]pyrrole-1-carboxamide;N-(1-(cyclopropylamino)-1,2-dioxohexan-3-yl)-2((S)-3-methyl-2-(piperidin-1-yl)butanoyl)octahydrocyclopenta[c]pyrrole-1-carboxamide; ethyl1-((2S,4R)-1((S)-3,3-dimethyl-2-(piperidin-1-yl)butanoyl)-4-(7-methoxy-2-phenylquinolin-4-yloxy)pyrrolidine-2-carboxamido)-2-vinylcyclopropanecarboxylate;ethyl1-((2S,4R)-4-(7-methoxy-2-phenylquinolin-4-yloxy)-1((S)-2-phenyl-2-(piperidin-1-yl)acetyl)pyrrolidine-2-carboxamido)-2-vinylcyclopropanecarboxylate;ethyl1-((2S,4R)-4-(7-methoxy-2-phenylquinolin-4-yloxy)-1((S)-3-methyl-2-(piperidin-1-yl)butanoyl)pyrrolidine-2-carboxamido)-2-vinylcyclopropanecarboxylate;ethyl1-((2S,4R)-1((S)-2-cyclohexyl-2-(piperidin-1-yl)acetyl)-4-(7-methoxy-2-phenylquinolin-4-yloxy)pyrrolidine-2-carboxamido)-2-vinylcyclopropanecarboxylate;ethyl1-((2S,4R)-4-(7-methoxy-2-phenylquinolin-4-yloxy)-1-((S)-2-(piperidin-1-yl)propanoyl)pyrrolidine-2-carboxamido)-2-vinylcyclopropanecarboxylate;1-((2S,4R)-1((S)-3,3-dimethyl-2-(piperidin-1-yl)butanoyl)-4-(7-methoxy-2-phenylquinolin-4-yloxy)pyrrolidine-2-carboxamido)-2-vinylcyclopropanecarboxylicacid;1-((2S,4R)-4-(7-methoxy-2-phenylquinolin-4-yloxy)-1((S)-2-phenyl-2-(piperidin-1-yl)acetyl)pyrrolidine-2-carboxamido)-2-vinylcyclopropanecarboxylicacid;1-((2S,4R)-4-(7-methoxy-2-phenylquinolin-4-yloxy)-1((S)-3-methyl-2-(piperidin-1-yl)butanoyl)pyrrolidine-2-carboxamido)-2-vinylcyclopropanecarboxylicacid; 1-((2S,4R)-1-((S)-2-cyclohexyl-2-(piperidin-1-yl)acetyl)-4-(7-methoxy-2-phenylquinolin-4-yloxy)pyrrolidine-2-carboxamido)-2-vinylcyclopropanecarboxylicacid;1-((2S,4R)-4-(7-methoxy-2-phenylquinolin-4-yloxy)-1-((S)-2-(piperidin-1-yl)propanoyl)pyrrolidine-2-carboxamido)-2-vinylcyclopropanecarboxylicacid; (2R,6S,16aS,Z)-ethyl2-(7-methoxy-2-phenylquinolin-4-yloxy)-5,16-dioxo-6-(piperidin-1-yl)-1,2,3,5,6,7,8,9,10,11,13a,14,14a,15,16,16a-hexadecahydrocyclopropa[e]pyrrolo[1,2-a][1,4]diazacyclopentadecine-14a-carboxylate;(2R,6S,16aS,Z)-2-(7-methoxy-2-phenylquinolin-4-yloxy)-5,16-dioxo-6-(piperidin-1-yl)-1,2,3,5,6,7,8,9,10,11,13a,14,14a,15,16,16a-hexadecahydrocyclopropa[e]pyrrolo[1,2-a][1,4]diazacyclopentadecine-14a-carboxylicacid; (2R,6S,13aR,14aS,16aS,Z)-ethyl2-(7-methoxy-2-phenylquinolin-4-yloxy)-5,16-dioxo-6-(piperidin-1-yl)-1,2,3,5,6,7,8,9,10,11,13a,14,14a,15,16,16a-hexadecahydrocyclopropa[e]pyrrolo[1,2-a][1,4]diazacyclopentadecine-14a-carboxylate;(2R,6S,13aS,14aR,16aS,Z)-ethyl2-(7-methoxy-2-phenylquinolin-4-yloxy)-5,16-dioxo-6-(piperidin-1-yl)-1,2,3,5,6,7,8,9,10,11,13a,14,14a,15,16,16a-hexadecahydrocyclopropa[e]pyrrolo[1,2-a][1,4]diazacyclopentadecine-14a-carboxylate;(2S,4R)-1-((S)-2-cyclohexyl-2-(piperidin-1-yl)acetyl)-N-(1-(cyclopropylsulfonylcarbamoyl)-2-vinylcyclopropyl)-4-(7-methoxy-2-phenylquinolin-4-yloxy)pyrrolidine-2-carboxamide;(2S,4R)-N-((1S,2R)-1-(cyclopropylsulfonylcarbamoyl)-2-vinylcyclopropyl)-4-(7-methoxy-2-phenylquinolin-4-yloxy)-1-((S)-2-phenyl-2-(piperidin-1-yl)acetyl)pyrrolidine-2-carboxamide;(2S,4R)-N-((1R,2S)-1-(cyclopropylsulfonylcarbamoyl)-2-vinylcyclopropyl)-4-(7-methoxy-2-phenylquinolin-4-yloxy)-1-((S)-2-phenyl-2-(piperidin-1-yl)acetyl)pyrrolidine-2-carboxamide;(2S,4R)-N-(1-(cyclopropylsulfonylcarbamoyl)-2-vinylcyclopropyl)-4-(7-methoxy-2-phenylquinolin-4-yloxy)-1-((S)-2-(piperidin-1-yl)propanoyl)pyrrolidine-2-carboxamide;(2S,4R)-N-(1-(cyclopropylsulfonylcarbamoyl)-2-vinylcyclopropyl)-4-(7-methoxy-2-phenylquinolin-4-yloxy)-1-((S)-3-methyl-2-(piperidin-1-yl)butanoylpyrrolidine-2-carboxamide;or(2S,4R)-N-(1-(cyclopropylsulfonylcarbamoyl)-2-vinylcyclopropyl)-4-(7-methoxy-2-phenylquinolin-4-yloxy)-1-((S)-4-methyl-2-(piperidin-1-yl)pentanoyl)pyrrolidine-2-carboxamide.